CN117897475A - Development of Yeast strains for ethanol production - Google Patents
Development of Yeast strains for ethanol production Download PDFInfo
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- CN117897475A CN117897475A CN202280038858.XA CN202280038858A CN117897475A CN 117897475 A CN117897475 A CN 117897475A CN 202280038858 A CN202280038858 A CN 202280038858A CN 117897475 A CN117897475 A CN 117897475A
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- yeast
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- saccharomyces
- fermentation
- ethanol
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Classifications
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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Abstract
Disclosed herein are yeast strains and derivatives thereof, and compositions comprising yeast strains for ethanol production. The present disclosure also relates to methods of producing ethanol from biomass using yeast strains and compositions. In particular, yeast strains produce lower glycerol and higher ethanol than the strains and products currently used in ethanol production processes, and have higher temperature tolerance and higher fermentation rates.
Description
Cross Reference to Related Applications
This patent application claims priority from U.S. provisional application No. 63/170,884 filed on 5, 4, 2021, the entire contents of which are incorporated herein by reference.
Technical Field
The present disclosure relates to yeast strains and derivatives thereof, and compositions comprising yeast strains for ethanol production. The present disclosure also relates to methods of producing ethanol from biomass using yeast strains and compositions. In particular, yeast strains produce lower glycerol and higher ethanol than the strains and products currently used in ethanol production processes, and have higher temperature and organic acid tolerance and higher fermentation rates.
Background
Ethanol may be produced from organisms using different biochemical pathways inherent to biological organisms. Ethanol produced by biological organisms is known as bioethanol and is therefore distinguished from ethanol produced by pure methods. Bioethanol is commercially produced and can be used as a liquid fuel in internal combustion engines (fuel ethanol), as a component in industrial products (industrial ethanol), or as a component in alcoholic beverages (potable ethanol).
Bioethanol is produced by biological organisms such as yeast. The yeast selected is advantageous for ethanol production, so that the production process and profitability can be optimized. Yeasts for commercial ethanol production require several characteristics, including adequate ethanol metabolic yield, adequate ethanol tolerance, acceptable byproduct yield, adequate fermentation kinetics, and a degree of heat resistance during fermentation. Yeasts of the genus Saccharomyces exhibit many of these characteristics that are required for commercial ethanol production. Commercial yeast products comprising yeast (Saccharomyces) include: ethanol Angel Super/>And Fali (AB->). Despite the favorable characteristics of commercial yeast strains and products for the production of ethanol, there is an increasing need to increase the efficiency of ethanol production to reduce the cost of manufacture.
Thus, there is a need for new and improved yeast (Saccharomyces) strains from which new and improved yeast products are produced. Improved yeast strains and products are needed to be able to increase the efficiency of commercial ethanol production by providing higher sugar to ethanol conversion, higher temperature tolerance, higher fermentation inhibitor tolerance and faster fermentation kinetics than the yeast strains and products currently in commercial use.
Disclosure of Invention
In one aspect, the disclosure relates to a yeast (Saccharomyces) strain selected from the group consisting of: (a) Yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003; (b) Yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004; (c) Yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005; (d) Yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006; (e) Yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit No. Y-68007; (f) Yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009.
In another aspect, the disclosure relates to a derivative of a yeast (Saccharomyces) strain selected from the group consisting of: (a) Yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003; (b) Yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004; (c) Yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005; (d) Yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006; (e) Yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit No. Y-68007; (f) Yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009. In one embodiment, the yeast strain or the derivative comprises one or more defined features selected from the group consisting of: (a) Under the same fermentation conditions, higher ethanol yields than yeast (Saccharomyces) strain Y1609; (b) Has increased temperature tolerance compared to yeast (Saccharomyces) strain Y1609 at a temperature of 32℃to 36 ℃; (c) Lower glycerol yields than yeast (Saccharomyces) strain Y1609 under the same fermentation conditions; (d) Has increased organic acid tolerance compared to yeast (Saccharomyces) strain Y1609 in the presence of an organic acid at a pH of from about 4.0 to about 5.2; and (e) has an increased fermentation rate compared to yeast (Saccharomyces) strain Y1609 under the same fermentation conditions. In another embodiment, the yeast strain or the derivative produces at least about 0.5% more ethanol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609. In another embodiment, the yeast strain or the derivative produces at least about 2% less glycerol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609. In another embodiment, the yeast strain or the derivative has a fermentation rate after 24 hours of fermentation that is at least about 2% greater than that of saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609. In another embodiment, the temperature is 32 ℃. In another embodiment, the yeast strain or the derivative produces at least about 5% more ethanol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609. In another embodiment, the yeast strain or the derivative produces at least about 3% less glycerol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609. In another embodiment, the yeast strain or the derivative has a fermentation rate after 24 hours of fermentation that is at least about 10% higher than that of saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609. In another embodiment, the temperature is 36 ℃. In another embodiment, the organic acid comprises lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, or a combination thereof.
Another aspect of the present disclosure provides a method of producing a derivative of a yeast (Saccharomyces) strain as described herein, comprising: (a) providing: (i) A first yeast strain, wherein the first yeast strain is selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; and (ii) a second yeast strain, wherein the second yeast strain is in a yeast narrow strain (Saccharomyces sensu stricto) clade; (b) Inducing sporulation of the first yeast strain and the second yeast strain; (c) Screening and selecting spores from the first yeast strain and spores from the second yeast strain; (d) Hybridizing selected spores of the first yeast strain with selected spores of the second yeast strain; and (e) screening or selecting the derivative strain. In one embodiment, step (c) comprises screening or selecting spores that exhibit one or more defined characteristics of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof. In another embodiment, step (e) comprises screening or selecting hybrids that exhibit one or more defined characteristics of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.
Another aspect of the present disclosure provides a method of producing a derivative of a yeast (Saccharomyces) strain as described herein, comprising: (a) providing: (i) A first yeast strain, wherein the first yeast strain is selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; and (ii) one or more additional yeast strains in the yeast narrow (Saccharomyces sensu stricto) clade; (b) Inducing sporulation of the first yeast strain and the one or more additional yeast strains to produce spores; (c) Mixing all spores of step (b) to allow hybridization of the spores; and (d) screening or selecting the derivative strain. In one embodiment, step (d) comprises screening or selecting hybrids that exhibit one or more defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929.
Another aspect of the present disclosure provides a mutant yeast of a yeast strain as described herein or a derivative as described herein.
Another aspect of the present disclosure provides a method of producing a mutant yeast as described herein, wherein the mutant yeast is mutated by contacting the yeast strain with a mutagen. In one embodiment, the mutagen is Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine 2 (ICR-170), or nitrogen mustard.
Another aspect of the present disclosure provides a method of producing a mutant yeast as described herein, wherein the mutant yeast is mutated by contacting the derivative with a mutagen. In one embodiment, the mutagen is Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine 2 (ICR-170), or nitrogen mustard.
Another aspect of the present disclosure provides an evolved yeast of a yeast strain as described herein or a derivative as described herein.
Another aspect of the present disclosure provides a method of producing an evolved yeast as described herein, wherein the evolution is induced by applying a selection pressure to the yeast strain.
Another aspect of the present disclosure provides a method of producing an evolved yeast as described herein, wherein the evolution is induced by applying a selection pressure to the derivative.
Another aspect of the present disclosure provides a genetically modified yeast of a yeast strain as described herein or a derivative as described herein. In one embodiment, the nucleic acid sequence of the genetically modified yeast is altered using genetic editing.
Another aspect of the present disclosure provides a recombinant yeast of a yeast strain as described herein or a derivative as described herein. In one embodiment, the recombinant yeast comprises modifications to inhibit gene expression, enhance gene expression, introduce genes, or delete genes.
Another aspect of the present disclosure provides a method of producing ethanol from a substrate by contacting the substrate with a fermenting organism, wherein the fermenting organism is selected from the group consisting of: (a) A yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003, or a derivative thereof; (b) A yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004, or a derivative thereof; (c) A yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005, or a derivative thereof; (d) A yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006, or a derivative thereof; (e) A yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit number Y-68007, or a derivative thereof; (f) A yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008, or a derivative thereof; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009, or a derivative thereof. In one embodiment, the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or a combination thereof. In another embodiment, the yeast strain comprises one or more defined features selected from the group consisting of: (a) Under the same fermentation conditions, higher ethanol yields than yeast (Saccharomyces) strain Y1609; (b) Has increased temperature tolerance compared to yeast (Saccharomyces) strain Y1609 at a temperature of 32℃to 36 ℃; (c) Lower glycerol yields than yeast (Saccharomyces) strain Y1609 under the same fermentation conditions; (d) Has increased organic acid tolerance compared to yeast (Saccharomyces) strain Y1609 in the presence of an organic acid at a pH of from about 4.0 to about 5.2; and (e) has an increased fermentation rate compared to yeast (Saccharomyces) strain Y1609 under the same fermentation conditions. In another embodiment, the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof. In another embodiment, the ethanol is produced using starch. In another embodiment, simultaneous Saccharification and Fermentation (SSF) or continuous fermentation is performed to produce the ethanol. In another embodiment, the ethanol is produced using a sugar. In another embodiment, batch fermentation or continuous fermentation is used to produce the ethanol. In another embodiment, the ethanol is produced using lignocellulosic sugars. In another embodiment, simultaneous Saccharification and Fermentation (SSF) or Step Hydrolysis Fermentation (SHF) is used to produce the ethanol.
Another aspect of the present disclosure provides a composition comprising a yeast strain as described herein or a derivative as described herein and one or more components selected from the group consisting of surfactants, emulsifiers, gums, swelling agents, protectants, and antioxidants. In one embodiment, the composition comprises one or more defined features selected from the group consisting of: (a) Higher ethanol yields than Fali under the same fermentation conditions; (b) Has increased temperature tolerance compared to Fali at temperatures of 32 ℃ to 36 ℃; (c) Lower glycerol yields than Fali under the same fermentation conditions; (d) Having increased organic acid tolerance compared to Fali at a pH of from about 4.0 to about 5.2 in the presence of an organic acid; and (e) has an increased fermentation rate compared to Fali under the same fermentation conditions.
Another aspect of the present disclosure provides a method of producing ethanol from biomass by contacting the biomass with a composition as described herein. In one embodiment, the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof. In another embodiment, the ethanol is produced using starch. In another embodiment, simultaneous Saccharification and Fermentation (SSF) or continuous fermentation is performed to produce the ethanol. In another embodiment, the ethanol is produced using a sugar. In another embodiment, batch fermentation or continuous fermentation is used to produce the ethanol. In another embodiment, the ethanol is produced using lignocellulosic sugars. In another embodiment, simultaneous Saccharification and Fermentation (SSF) or Step Hydrolysis Fermentation (SHF) is used to produce the ethanol.
Another aspect of the present disclosure provides a method of producing a fermentation product from a substrate by contacting the substrate with a fermenting organism, wherein the fermenting organism is selected from the group consisting of: (a) A yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003, or a derivative thereof; (b) A yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004, or a derivative thereof; (c) A yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005, or a derivative thereof; (d) A yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006, or a derivative thereof; (e) A yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit number Y-68007, or a derivative thereof; (f) A yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008, or a derivative thereof; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009, or a derivative thereof. In one embodiment, the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or a combination thereof. In another embodiment, the fermentation product is ethanol. In another embodiment, the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof. In another embodiment, the fermentation product is produced using batch fermentation, continuous fermentation, simultaneous Saccharification and Fermentation (SSF), or Step Hydrolysis Fermentation (SHF).
The present disclosure provides other aspects and embodiments that will be apparent from the following detailed description and the accompanying drawings.
Drawings
FIG. 1 shows the process of making a new yeast strain. Fig. 1A shows directional mating. Fig. 1B shows a large-scale mating. FIG. 1C shows directed evolution of yeast strains. FIG. 1D shows the mutation of the yeast strain.
Figure 2 shows the percentage difference of no exogenously added organic acid relative to control (Y1609) at the end of 50 hours fermentation at normal fermentation temperature (32 ℃) in proprietary synthetic corn starch medium (SCM) at pH 5.2. The percentage differences of ethanol at the end of fermentation (fig. 2A), glycerol at the end of fermentation (fig. 2B) and the initial fermentation rate of fermentation (fig. 2C) relative to the control are shown.
FIG. 3 shows the percentage difference of exogenously added 1% w/v lactic acid and 0.05% w/v acetic acid relative to the control at the end of 50 hours fermentation at normal fermentation temperature (32 ℃) in proprietary synthetic corn starch medium (SCM) at pH 4.0. The percentage differences of ethanol at the end of fermentation (fig. 3A), glycerol at the end of fermentation (fig. 3B) and the initial fermentation rate of fermentation (fig. 3C) relative to the control are shown.
Figure 4 shows the percentage difference of no exogenously added organic acid relative to control (Y1609) at the end of 50 hours fermentation at high fermentation temperature (36 ℃) in proprietary synthetic corn starch medium (SCM) at pH 5.2. The percentage differences of ethanol at the end of fermentation (fig. 4A), glycerol at the end of fermentation (fig. 4B) and the initial fermentation rate of fermentation (fig. 4C) relative to the control are shown.
FIG. 5 shows the percentage difference of exogenously added 1% w/v lactic acid and 0.05% w/v acetic acid relative to control (Y1609) at the end of 50 hours fermentation at high fermentation temperature (36 ℃) in proprietary synthetic corn starch medium (SCM) at pH 4.0. The percentage difference of ethanol at the end of fermentation (fig. 5A), glycerol at the end of fermentation (fig. 5B) and the initial fermentation rate of fermentation (fig. 5C) relative to the control is shown.
Detailed Description
Described herein are fermenting organisms, including current industry standard yeasts used in yeast products (e.g., fali) that have defined characteristics including higher ethanol production, lower glycerol production, higher temperature and inhibitor tolerance, and higher fermentation rates, under the same fermentation conditions. Also described herein are yeast (Saccharomyces) strains having improved properties compared to the yeast strains used in Fali. The present disclosure relates to methods of producing yeast products from yeast strains. The present disclosure also relates to improved methods of producing ethanol from different fermentable biomass materials using the fermenting organisms described herein.
1. Definition of the definition
Unless defined otherwise, all technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art. In the event of a conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.
As used herein, the terms "comprising," "including," "having," "can," "containing," and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not exclude additional behavioral or structural possibilities. The singular forms "a", "and" the "include plural referents unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments "comprising," "consisting of … …," and "consisting essentially of … …," embodiments or elements described herein, whether or not explicitly stated.
To describe the numerical ranges herein, each intermediate number having the same degree of precision is expressly contemplated. For example, for a range of 6-9, the numbers 7 and 8 are also contemplated in addition to 6 and 9; for the range of 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 and 7.0 are explicitly contemplated; and for a range of 1 to 5, the numbers 2, 3 and 4 are considered in addition to 1 and 5.
The term "about" or "approximately" as used herein for one or more values of interest refers to a value that is similar to a specified reference value, or a value that is within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on the manner in which the value is measured or determined, such as the limitations of the measurement system. In certain aspects, the term "about" refers to a range of values that fall within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater than or less than) of a specified reference value, unless otherwise stated or the context indicates otherwise (unless the number exceeds 100% of the possible values). Alternatively, "about" may mean within 3 or more standard deviations, according to practice in the art. Alternatively, for example with respect to biological systems or processes, the term "about" may mean within an order of magnitude, preferably within 5 times a value, and more preferably within 2 times a value.
As used herein, the term "biomass" refers to any organic matter of plant origin that can become a source of carbohydrates after conversion. Preferably, the biomass may be from agricultural or food processing products and/or byproducts. In particular, the biomass may be enriched in sucrose or starch and is selected from or derived from, for example, sorghum, sugarcane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca or mixtures thereof.
As used herein, "combination of DNA" between yeast strains refers to a combination of the entire genome or a portion of the genome of a yeast strain. The combination of DNA between yeast strains may be by any method suitable for combining DNA of at least two yeast cells, and may include, for example, mating methods that include sporulation of yeast strains to produce haploid cells and subsequent crossing or mating compatible haploid cells; cell induction; or cell fusion, such as protoplast fusion.
As used herein, a "derivative" is a yeast strain derived from a yeast strain disclosed herein (e.g., saccharomyces (Saccharomyces) or a clade of Saccharomyces narrow (Saccharomyces sensu stricto)), including by hybridization, mutagenesis, recombinant DNA techniques, mating, cell fusion, or cell induction between yeast strains. The derivative strain may be the direct progeny (i.e., the product of mating between the strain of the invention and another strain or itself).
As used herein, "ethanol yield of glucose" is the yield of ethanol obtained from glucose. In one embodiment, the ethanol yield of glucose is represented by the following description: one molecule of glucose produces two molecules of ethanol and two molecules of carbon dioxide. In another embodiment, the ethanol yield of glucose is represented by the formula: c (C) 6 H 12 O 6 →2C 2 H 5 OH+2CO 2 Wherein C 6 H 12 O 6 Is of the formula glucose, C 2 H 5 OH is the chemical formula of ethanol and CO 2 Is the chemical formula of carbon dioxide. In another embodiment, the ethanol production of glucose is expressed on a mass basis, wherein 1.0 gram of glucose produces 0.511 grams of ethanol and 0.489 grams of carbon dioxide. The highest ethanol yield of glucose is one molecule of glucose producing two molecules of ethanol. The highest ethanol yield on a mass basis is 0.511 gram ethanol from one gram glucose.
The terms "control" and "Y1609" are used interchangeably when discussing yeast strains, and the terms "control" and "Fali" are used interchangeably when discussing yeast products. Strain Y1609 was used to produce the yeast product Fali. Fali can be produced as active dry yeast products, crushed yeast products, and liquid yeast products for fermenting substrates to produce fuel ethanol, industrial ethanol, and potable ethanol. Fali is particularly useful for the fermentation of these sugars released from sucrose, glucose and fructose-containing biomass, as well as the fermentation of sugars released from starch-containing biomass after release of the sugars by enzymatic or chemical processes. Fali can be used for starch bottom Batch fermentation, continuous fermentation, and Simultaneous Saccharification and Fermentation (SSF) of the material. It has high tolerance to released glucose and high cell count>20x10 9 Individual cells/g), medium ethanol and temperature tolerance, medium organic acid tolerance. It is well rehydrated in direct delivery (pitch) applications and can be used with saccharification enzymes and alpha-amylase enzyme systems. Fali has optimal performance in the pH range of 4.0 to 5.0, but ferments well in the pH range of 3.5 to 6.0. The optimal fermentation temperature for Fali depends on the pressure present (e.g., organic acid, ethanol, and pH), but typically ferments well over a temperature range of about 32 ℃. Fali can be obtained from ABAre commercially available.
The term "fermentation medium" refers to an environment in which fermentation is performed using a fermenting organism, including fermentable substrates, i.e., a carbohydrate source (e.g., glucose) that is metabolized by the fermenting organism into a desired fermentation product (e.g., ethanol). The fermentation medium may comprise fermentation nutrients of the fermenting organism. Fermentation nutrients are widely used in fermentation technology and include nitrogen sources (such as urea ammonia, vitamins, minerals, or combinations thereof). "feed" is a fermentation medium, however, the composition of the feed may be different from the fermentation medium.
As used herein, "high yield ethanol production" refers to the production of ethanol by fermentation, wherein the ethanol yield approximates that of glucose. High ethanol production requires limited by-products (e.g., glycerol) and yeast growth during fermentation.
As used herein, the terms "enhanced," "increased," "enhanced," or "larger" refer to an increase or improvement in a particular characteristic or trait as compared to other similar organisms, control or wild-type organisms. Typically, this is a beneficial trait associated with fermentation.
The term "inoculum" means the amount of microorganisms added to the main fermenter to start the fermentation process. In fermentation processes using seed fermenters, the inoculum is typically an amount of preculture corresponding to 5% to 20% of the volume of the main fermentor.
The term "isolated" refers to a form or an environment in which matter does not exist in nature. Non-limiting examples of isolated substances include (1) any non-naturally occurring substance, (2) any substance, including but not limited to any enzyme, variant, nucleic acid, protein, peptide, or cofactor, that is at least partially removed from one or more or all of the naturally occurring components with which it is associated in nature; (3) Any substance that is modified by humans compared to substances found in nature; or (4) any substance modified by increasing the amount of the substance relative to other components with which it is naturally associated (e.g., recombinant production in yeast; multiple copies of a gene encoding the substance; and use of a stronger promoter than the promoter naturally associated with the gene encoding the substance). The separated material may be present in the fermentation broth sample; for example, yeast may be genetically modified to express specific polypeptides. The fermentation broth from the yeast will comprise the isolated polypeptide.
The term "low pH" as used herein refers to a pH of about 2.5 to about 4.5. The low pH is preferably less than about 4.5. As used herein, the term "normal pH" refers to a pH of about 4.0 to about 6.0. The normal pH is preferably about 5.0.
The term "main fermenter" as used herein is the final fermenter used in a fermentation process for producing a fermentation product, wherein the desired fermentation product is produced.
The term "parent" or "parent" strain refers to a yeast strain from which the derivative strain is derived. In some embodiments, the derivative may also be the parent.
The term "preculture" is understood as a liquid positive growth culture for inoculating microorganisms (i.e. yeasts) of the main fermenter. Positive growth means that the culture is in a stage where the microorganism increases the number of cells. Precultures are often used as seeding materials to avoid or reduce lag phases in the main fermenter. During fermentation to produce ethanol, the cells in the pre-fermentor are typically conditioned. The idea is not to produce yeast biomass because the carbon in the biomass reduces the carbon entering the ethanol. There is fermentative growth, but it is undesirable to promote aerobic growth (e.g., using a seed fermenter). Alternatively, yeast may be added directly to the main fermenter by "direct feeding".
As used herein, the terms "property" and "defined characteristics" of the saccharomyces cerevisiae (Saccharomyces cerevisiae) strain detailed herein include at least increased ethanol production under the same process conditions as compared to a control (i.e., fali or Y1609). Other "properties" and "defined characteristics" include, inter alia, increased temperature tolerance, increased fermentation rate, increased tolerance to organic acids, increased ethanol production, and decreased glycerol production. For example, a fermenting organism described herein used in a process described herein can have one or more of the above "properties" and "defining characteristics.
The term "pre-fermenter" means a fermenter in which a pre-culture is formed by fermenting microorganisms until yeast is activated and conditioned for inoculation into the main fermenter. When the pre-fermenter is not used, a "direct launch" is used.
As used herein, a "substrate" is a molecule that can be directly or indirectly metabolized to ethanol by fermentation of yeast (Saccharomyces) or any of the yeasts or yeast products described herein.
The term "wild type" as used herein refers to a typical form of an organism or genetic material thereof, as it typically occurs, as distinguished from a selected organism.
The terms "yeast product" and "composition" are used interchangeably herein and as used herein refer to a composition comprising, among other things, dry yeast, starch, and an emulsifier. The yeast product may also be a liquid composition comprising, among other things, cream yeast, glycerin and xanthan gum.
Unless defined otherwise herein, scientific and technical terms related to the present disclosure shall have the meanings commonly understood by one of ordinary skill in the art. For example, any terms and techniques described herein relating to cell biology, molecular biology, microbiology, genetics, protein and nucleic acid chemistry are well known and commonly used in the art. The meaning and scope of the terms should be clear; however, if there are any potential ambiguities, the definitions provided herein take precedence over any dictionary or extrinsic definitions. Furthermore, unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular.
2. Yeast strain and yeast strain derivative
As used herein, a yeast strain and yeast strain derivative may be any yeast used in ethanol production, including, but not limited to, saccharomyces (Saccharomyces), zygoSaccharomyces (zygo Saccharomyces), saccharomyces Brettanomyces (Brettanomyces) and Kluyveromyces (Kluyveromyces). Preferably, the yeast may be Saccharomyces cerevisiae (Saccharomyces sp.) and even more preferably it may be Saccharomyces cerevisiae (Saccharomyces cerevisiae).
Furthermore, the yeast (Saccharomyces) strains and derivatives thereof described herein can be readily distinguished from the following: (a) a naturally occurring strain of yeast (Saccharomyces); (b) a contaminated yeast (Saccharomyces) strain; and (c) other strains used in the ethanol industry that do not have ethanol production capacity and the defined characteristics of the strains described herein.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher ethanol yield than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher ethanol yield at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher ethanol yield at a temperature of 32 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In another specific embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher ethanol yield at a temperature of 36 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. The inventors surprisingly found that the yeast strains described herein lead to statistically significantly higher ethanol yields compared to Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein generally lead to statistically significantly higher ethanol yields (e.g., Y1919 in fig. 2-5) than Y1609 under the same conditions.
In some embodiments, the yeast strains and derivatives thereof as described herein produce at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% more ethanol after 50 hours of fermentation relative to a typical yeast strain used for fermentation (e.g., yeast (Saccharomyces) strain Y1609). In some embodiments, the yeast strains and derivatives thereof as described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% ethanol after 50 hours of fermentation relative to a typical yeast strain used for fermentation (e.g., yeast (Saccharomyces) strain Y1609). In some embodiments, the yeast strain is selected from the group consisting of yeast (Saccharomyces) strain Y1609, the yeast strains and derivatives thereof as described herein produce about 0.1% -21% more (i.e., from about 0.1% to about 21%), 0.2% -21%, 0.3% -21%, 0.4% -21%, 0.5% -21%, 0.6% -21%, 0.7% -21%, 0.8% -21%, 0.9% -21%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 0.2% -20%, 0.3% -20%, 0.4% -20%, 0.5% -20%, 0.6% -20% after 50 hours of fermentation 0.6% -20%, 0.7% -20%, 0.8% -20%, 0.9% -20%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 0.2% -19%, 0.3% -19%, 0.4% -19%, 0.5% -19%, 0.6% -19%, 0.7% -19%, 0.8% -19%, 0.9% -19%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 5% -19%, and the like, 10% -19%, 0.2% -18%, 0.3% -18%, 0.4% -18%, 0.5% -18%, 0.6% -18%, 0.7% -18%, 0.8% -18%, 0.9% -18%, 1% -18%, 2% -18%, 3% -18%, 4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18%, 9% -18%, 10% -18%, 0.2% -17%, 0.3% -17%, 0.4% -17%, 0.5% -17%, 0.6% -17%, 0.7% -17%, 0.8% -17%, 0.9% -17%, 1% -17%, 2% -17%, 3% -17%, 4% -17%, 5% -17%, 6% -17% and the like 7% -17%, 8% -17%, 9% -17%, 10% -17%, 0.2% -16%, 0.3% -16%, 0.4% -16%, 0.5% -16%, 0.6% -16%, 0.7% -16%, 0.8% -16%, 0.9% -16%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, 5% -16%, 6% -16%, 7% -16%, 8% -16%, 9% -16%, 10% -16%, 0.2% -15%, 0.3% -15%, 0.4% -15%, 0.5% -15%, 0.6% -15%, 0.7% -15%, 0.8% -15%, 0.9% -15%, 1% -15%, 2% -15% and, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 0.1% -14%, 0.2% -14%, 0.3% -14%, 0.4% -14%, 0.5% -14%, 0.6% -14%, 0.7% -14%, 0.8% -14%, 0.9% -14%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 0.1% -13%, 0.2% -13%, 0.3% -13%, 0.4% -13%, 0.5% -13%, 0.6% -13%, 0.8% -14%, 1% -14%, 2% -14%, 5% -14%, 6% -13% and 0.3% -13% of the composition 0.7% -13%, 0.8% -13%, 0.9% -13%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 0.1% -12%, 0.2% -12%, 0.3% -12%, 0.4% -12%, 0.5% -12%, 0.6% -12%, 0.7% -12%, 0.8% -12%, 0.9% -12%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 0.2% -12%, 0.8% -12%, 0.9% -12%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 0.1% -11%, 0.2% -11%, 0.3% -11%, 0.4% -11%, 0.5% -11%, 0.6% -11%, 0.7% -11%, 0.8% -11%, 0.9% -11%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 0.1% -0.2%, 0.1% -0.3%, 0.1% -0.4%, 0.1% -0.5%, 0.1% -0.6%, 0.1% -0.7%, 0.1% -0.8%, 0.1% -0.9%, 0.1% -1%, 0.1% -2%, 0.1% -3%, 0.1% -4%, 0.1% -5%, 0.1% -6%, 0.1% -7%, 0.1% -8% or 0.1% -9% ethanol.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly lower glycerol yield than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly lower glycerol yield at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly lower glycerol yield at a temperature of 32 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In another particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly lower glycerol yield at a temperature of 36 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. The inventors surprisingly found that the yeast strains described herein lead to statistically significantly lower glycerol yields compared to Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein lead to statistically significantly lower glycerol yields (e.g., Y1919 in fig. 2-5) than Y1609 under the same conditions.
In some embodiments, a yeast strain as described herein and derivatives thereof produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, or 22% less glycerol after 50 hours of fermentation relative to a typical yeast strain used for fermentation, e.g., yeast (Saccharomyces) strain Y1609. In some embodiments, a yeast strain as described herein and derivatives thereof produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% less glycerol after 50 hours of fermentation relative to a typical yeast strain used for fermentation, e.g., yeast (Saccharomyces) strain Y1609. In some embodiments, the yeast strains and derivatives thereof as described herein produce about 1% -30% (i.e., from about 1% -30%), 2% -30%, 3% -30%, 4% -30%, 5% -30%, 6% -30%, 7% -30%, 8% -30%, 9% -30%, 10% -30%, 11% -30%, 12% -30%, 13% -30%, 14% -30%, 15% -30%, 16% -30%, 17% -30%, 18% -30%, 19% -30%, 20% -30%, 21% -30%, 22% -30%, 23% -30%, 24% -30%, 25% -30%, 26% -30%, 27% -30%, 28% -30%, 29% -30%, 1% -29%, 2% -29%, 3% -29%, 4% -29%, 5% -29%, 6% -29%, 7% -29%, 8% -29%, 9% -29%, 10% -29%, 11% -29%, 12% -29%, 13% -29%, 14% -30%, 15% -30%, 17% -30%, 21% -30%, 22% -30%, 23% -30%, 24% -30%, 25% -26% -30%, 27% -30%, 29%, 7% -29%, 10% -29%, 11% -29%, 12% -29%, 13% -29%, 14% -29%, 15% -29%, 18% -29%, 21% -29%, 22% -29%, and the like after 50 hours of fermentation 26% -29%, 27% -29%, 28% -29%, 1% -28%, 2% -28%, 3% -28%, 4% -28%, 5% -28%, 6% -28%, 7% -28%, 8% -28%, 9% -28%, 10% -28%, 11% -28%, 12% -28%, 13% -28%, 14% -28%, 15% -28%, 16% -28%, 17% -28%, 18% -28%, 19% -28%, 20% -28%, 21% -28%, 22% -28%, 23% -28%, 24% -28%, 25% -28%, 26% -28%, 27% -28%, 1% -27%, 2% -27%, 3% -27%, and 4% -27%, 5% -27%, 6% -27%, 7% -27%, 8% -27%, 9% -27%, 10% -27%, 11% -27%, 12% -27%, 13% -27%, 14% -27%, 15% -27%, 16% -27%, 17% -27%, 18% -27%, 19% -27%, 20% -27%, 21% -27%, 22% -27%, 23% -27%, 24% -27%, 25% -27%, 26% -27%, 1% -26%, 2% -26, 3% -26%, 4% -26%, 5% -26%, 6% -26%, 7% -26%, 8% -26%, 9% -26%, 10% -26%, 11% -26%, 12% -26%, 13% -26%, 14% -26%, 15% -26%, 16% -26%, 17% -26%, 18% -26%, 19% -26%, 20% -26%, 21% -26%, 22% -26%, 23% -26%, 24% -26%, 25% -26%, 1% -25%, 2% -25%, 3% -25%, 4% -25%, 5% -25%, 6% -25%, 7% -25%, 8% -25%, 9% -25%, 10% -25%, 11% -25%, 12% -25%, 13% -25%, 14% -25%, 15% -25%, 16% -25%, 17% -25%, 18% -25%, 19% -25%, 20% -25%, 21% -25%, 22% -25%, 23% -25%, 24% -25%, 1% -24%, 2% -24%, 3% -24%, 4% -24%, 5% -24%, 6% -24%, 7% -24%, 8% -24%, 9% -24%, 10% -24%, 11% -24%, 12% -24%, 13% -24%, 14% -24%, 15% -24%, 16% -24%, 17% -24%, 18% -24%, 19% -24%, 20% -24%, 21% -24%, 22% -24%, 23% -24%, 1% -23%, 2% -23% and, 3% -23%, 4% -23%, 5% -23%, 6% -23%, 7% -23%, 8% -23%, 9% -23%, 10% -23%, 11% -23%, 12% -23%, 13% -23%, 14% -23%, 15% -23%, 16% -23%, 17% -23%, 18% -23%, 19% -23%, 20% -23%, 21% -23%, 22% -23%, 1% -22%, 2% -22%, 3% -22%, 4% -22%, 5% -22%, 6% -22%, 7% -22%, 8% -22%, 9% -22%, 10% -22%, 11% -22%, 12% -22%, 13% -22%, and 14% -22%, 15% -22%, 16% -22%, 17% -22%, 18% -22%, 19% -22%, 20% -22%, 21% -22%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 11% -21%, 12% -21%, 13% -21%, 14% -21%, 15% -21%, 16% -21%, 17% -21%, 18% -21%, 19% -21%, 20% -21%, 1% -20%, 2% -20%, 3% -20%, 4% -20% and, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 11% -20%, 12% -20%, 13% -20%, 14% -20%, 15% -20%, 16% -20%, 17% -20%, 18% -20%, 19% -20%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 10% -19%, 11% -19%, 12% -19%, 13% -19%, 14% -19%, 15% -19%, 16% -19%, 17% -19%, 18% -19%, and 1% -18%, 2% -18%, 3% -18%, 4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18%, 9% -18%, 10% -18%, 11% -18%, 12% -18%, 13% -18%, 14% -18%, 15% -18%, 16% -18%, 17% -18%, 1% -17%, 2% -17%, 3% -17%, 4% -17%, 5% -17%, 6% -17%, 7% -17%, 8% -17%, 9% -17%, 10% -17%, 11% -17%, 12% -17%, 13% -17%, 14% -17%, 15% -17% and, 16% -17%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, 5% -16%, 6% -16%, 7% -16%, 8% -16%, 9% -16%, 10% -16%, 11% -16%, 12% -16%, 13% -16%, 14% -16%, 15% -16%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 11% -15%, 12% -15%, 13% -15%, 14% -15%, 1% -14%, 2% -14%, 3% -14%, and 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 11% -14%, 12% -14%, 13% -14%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 11% -13%, 12% -13%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 11% -12%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 1% -10%, 2% -10%, 3% -10%, 4% -10%, 5% -10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -6%, 2% -6%, 3% -6%, 4% -6%, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -4%, 2% -4%, 1% -3%, 2% -3%, 3% -3% or 1% -2% glycerol.
In some embodiments, the yeast strains and derivatives thereof as described herein have a higher fermentation rate than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher fermentation rate at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher fermentation rate at a temperature of 32 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In another particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher fermentation rate at a temperature of 36 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. The inventors surprisingly found that under the same fermentation conditions, the yeast strains described herein caused a statistically significantly higher fermentation rate compared to Y1609. The inventors surprisingly found that the derivatives described herein lead to a statistically significantly higher fermentation rate (e.g., Y1919 in fig. 2-5) than Y1609 under the same conditions.
In some embodiments, the yeast strains and derivatives thereof as described herein have a fermentation rate after 24 hours of fermentation that is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% higher than a typical yeast strain used for fermentation, such as yeast (Saccharomyces) strain Y1609. In some embodiments, a yeast strain and derivatives thereof as described herein have a fermentation rate after 24 hours of fermentation that is up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% or 50% higher than a typical yeast strain used for fermentation, such as yeast (Saccharomyces) strain Y1609. In some embodiments, the yeast strains and derivatives thereof as described herein have a fermentation rate after 24 hours of fermentation that is about 1% -50% (i.e., from about 1% -50%, 3% -50%, 4% -50%, 5% -50%, 6% -50%, 7% -50%, 8% -50%, 9% -50%, 10% -50%, 11% -50%, 12% -50%, 13% -50%, 14% -50%, 15% -50%, 16% -50%, 17% -50%, 18% -50%, 19% -50%, 20% -50%, 21% -50%, 22% -50%, 23% -50%, 24% -50%, 25% -50%, 26% -50%, 27% -50%, 28% -50%, 2% -49%, 3% -49%, 4% -49%, 5% -49%, 6% -49%, 7% -49%, 8% -49%, 9% -49%, 10% -49%, 11% -49%, 12% -49%, 13% -49%, 14% -49%, 15% -49%, 16% -50%, 18% -50%, 19% -50%, 20% -50%, 22% -50%, 23% -50%, 24% -50%, 25% -50%, 26% -50%, 2% -49%, 4% -49%, 5% -49%, 6% -49%, 8% -49%, 10% -49%, 11% -49%, 12% -49%, 13% -49%, 14% -49%, 15% -49%, and 21% -49%, and 26% -49% of the typical yeast strains used for fermentation 28% -49%, 2% -48%, 3% -48%, 4% -48%, 5% -48%, 6% -48%, 7% -48%, 8% -48%, 9% -48%, 10% -48%, 11% -48%, 12% -48%, 13% -48%, 14% -48%, 15% -48%, 16% -48%, 17% -48%, 18% -48%, 19% -48%, 20% -48%, 21% -48%, 22% -48%, 23% -48%, 24% -48%, 25% -48%, 26% -48%, 27% -48%, 28% -48%, 2% -47%, 3% -47%, 4% -47%, 5% -47%, 6% -47%, and 7% -47%, 8% -47%, 9% -47%, 10% -47%, 11% -47%, 12% -47%, 13% -47%, 14% -47%, 15% -47%, 16% -47%, 17% -47%, 18% -47%, 19% -47%, 20% -47%, 21% -47%, 22% -47%, 23% -47%, 24% -47%, 25% -47%, 26% -47%, 27% -47%, 28% -47%, 2% -46%, 3% -46%, 4% -46%, 5% -46%, 6% -46%, 7% -46%, 8% -46%, 9% -46%, 10% -46%, 11% -46%, and, 12% -46%, 13% -46%, 14% -46%, 15% -46%, 16% -46%, 17% -46%, 18% -46%, 19% -46%, 20% -46%, 21% -46%, 22% -46%, 23% -46%, 24% -46%, 25% -46%, 26% -46%, 27% -46%, 28% -46%, 2% -45%, 3% -45%, 4% -45%, 5% -45%, 6% -45%, 7% -45%, 8% -45%, 9% -45%, 10% -45%, 11% -45%, 12% -45%, 13% -45%, 14% -45%, 15% -45%, 16% -45%, 17% -45%, 18% -45%, 19% -45%, 20% -45%, 21% -45%, 22% -45%, 23% -45%, 24% -45%, 25% -45%, 26% -45%, 27% -45%, 28% -45%, 2% -44%, 3% -44%, 4% -44%, 5% -44%, 6% -44%, 7% -44%, 8% -44%, 9% -44%, 10% -44%, 11% -44%, 12% -44%, 13% -44%, 14% -44%, 15% -44%, 16% -44%, 17% -44%, 18% -44%, 19% -44%, 20% -44%, 21% -44%, 22% -44%, 23% -44%, 24% -44%, 25% -44%, 26% -44%, 27% -44%, 28% -44%, 2% -43%, 3% -43%, 4% -43%, 5% -43%, 6% -43%, 7% -43%, 8% -43%, 9% -43%, 10% -43%, 11% -43%, 12% -43%, 13% -43%, 14% -43%, 15% -43%, 16% -43%, 17% -43%, 18% -43%, 19% -43%, 20% -43%, 21% -43%, 22% -43%, 23% -43%, 24% -43%, 25% -43%, 26% -43%, 27% -43%, 28% -43%, 2% -42%, 3% -42%, 4% -42%, 5% -42%, 6% -42%, 7% -42%, 8% -42%, 9% -42%, 10% -42%, 11% -42%, 12% -42%, 13% -42%, 14% -42%, 15% -42%, 16% -42%, 17% -42%, 18% -42%, 19% -42%, 20% -42%, 21% -42%, 22% -42%, 23% -42%, 24% -42%, 25% -42%, 26% -42%, 28% -42%, 2% -42%, 4% -41%, 5% -41%, and 4% -42% 7% -41%, 8% -41%, 9% -41%, 10% -41%, 11% -41%, 12% -41%, 13% -41%, 14% -41%, 15% -41%, 16% -41%, 17% -41%, 18% -41%, 19% -41%, 20% -41%, 21% -41%, 22% -41%, 23% -41%, 24% -41%, 25% -41%, 26% -41%, 27% -41%, 28% -41%, 2% -40%, 3% -40%, 4% -40%, 5% -40%, 6% -40%, 7% -40%, 8% -40%, 9% -40%, 10% -40%, 11% -40%, 12% -40%, and 13% -40%, 14% -40%, 15% -40%, 16% -40%, 17% -40%, 18% -40%, 19% -40%, 20% -40%, 21% -40%, 22% -40%, 23% -40%, 24% -40%, 25% -40%, 26% -40%, 27% -40%, 28% -40%, 2% -39%, 3% -39%, 4% -39%, 5% -39%, 6% -39%, 7% -39%, 8% -39%, 9% -39%, 10% -39%, 11% -39%, 12% -39%, 13% -39%, 14% -39%, 15% -39%, 16% -39%, 17% -39%, 18% -39%, 19% -39%, 20% -39%, 21% -39%, 22% -39%, 23% -39%, 24% -39%, 25% -39%, 26% -39%, 27% -39%, 28% -39%, 2% -38%, 3% -38%, 4% -38%, 5% -38%, 6% -38%, 7% -38%, 8% -38%, 9% -38%, 10% -38%, 11% -38%, 12% -38%, 13% -38%, 14% -38%, 15% -38%, 16% -38%, 17% -38%, 18% -38%, 19% -38%, 20% -38%, 21% -38%, 22% -38%, 23% -38%, and 24% -38%, 25% -38%, 26% -38%, 27% -38%, 28% -38%, 2% -37%, 3% -37%, 4% -37%, 5% -37%, 6% -37%, 7% -37%, 8% -37%, 9% -37%, 10% -37%, 11% -37%, 12% -37%, 13% -37%, 14% -37%, 15% -37%, 16% -37%, 17% -37%, 18% -37%, 19% -37%, 20% -37%, 21% -37%, 22% -37%, 23% -37%, 24% -37%, 25% -37%, 26% -37%, 27% -37%, 28% -37%, 2% -36%, 3% -36%, 4% -36%, 5% -36%, 6% -36%, 7% -36%, 8% -36%, 9% -36%, 10% -36%, 11% -36%, 12% -36%, 13% -36%, 14% -36%, 15% -36%, 16% -36%, 17% -36%, 18% -36%, 19% -36%, 20% -36%, 21% -36%, 22% -36%, 23% -36%, 24% -36%, 25% -36%, 26% -36%, 27% -36%, 28% -36%, 2% -35%, 3% -35%, 4% -35%, 5% -35%, 6% -35%, 7% -35%, 8% -35%, 9% -35%, 10% -35%, 11% -35%, 12% -35%, 13% -35%, 14% -35%, 15% -35%, 16% -35%, 17% -35%, 18% -35%, 19% -35%, 20% -35%, 21% -35%, 22% -35%, 23% -35%, 24% -35%, 25% -35%, 26% -35%, 27% -35%, 28% -35%, 2% -34%, 3% -34%, 4% -34%, 5% -35%, 6% -35%, 7% -35%, 34%, 8% -35%, 9% -35%, 10% -35%, 11% -35%, 12% -35%, 13% -35%, 14% -35%, 15% -35%, and 22% -35%, 23% -35%, 22% -35%, and 24% -35%, and the like 13% -34%, 14% -34%, 15% -34%, 16% -34%, 17% -34%, 18% -34%, 19% -34%, 20% -34%, 21% -34%, 22% -34%, 23% -34%, 24% -34%, 25% -34%, 26% -34%, 27% -34%, 28% -34%, 2% -33%, 3% -33%, 4% -33%, 5% -33%, 6% -33%, 7% -33%, 8% -33%, 9% -33%, 10% -33%, 11% -33%, 12% -33%, 13% -33%, 14% -33%, 15% -33%, 16% -33%, 17% -33%, 18% -33%, and 19% -33%, 20% -33%, 21% -33%, 22% -33%, 23% -33%, 24% -33%, 25% -33%, 26% -33%, 27% -33%, 28% -33%, 2% -32%, 3% -32%, 4% -32%, 5% -32%, 6% -32%, 7% -32%, 8% -32%, 9% -32%, 10% -32%, 11% -32%, 12% -32%, 13% -32%, 14% -32%, 15% -32%, 16% -32%, 17% -32%, 18% -32%, 19% -32%, 20% -32%, 21% -32%, 22% -32%, 23% -32%, 24% -32%, 25% -32%, 26% -32%, 27% -32%, 28% -32%, 2% -31%, 3% -31%, 4% -31%, 5% -31%, 6% -31%, 7% -31%, 8% -31%, 9% -31%, 10% -31%, 11% -31%, 12% -31%, 13% -31%, 14% -31%, 15% -31%, 16% -31%, 17% -31%, 18% -31%, 19% -31%, 20% -31%, 21% -31%, 22% -31%, 23% -31%, 24% -31%, 25% -31%, 26% -31%, 27% -31%, 28% -31%, 2% -30%, and 3% -30%, 4% -30%, 5% -30%, 6% -30%, 7% -30%, 8% -30%, 9% -30%, 10% -30%, 11% -30%, 12% -30%, 13% -30%, 14% -30%, 15% -30%, 16% -30%, 17% -30%, 18% -30%, 19% -30%, 20% -30%, 21% -30%, 22% -30%, 23% -30%, 24% -30%, 25% -30%, 26% -30%, 27% -30%, 28% -30%, 29% -30%, 1% -29%, 2% -29%, 3% -29%, 4% -29%, 6% -29%, 7% -29%, 8% -29%, 9% -29%, 10% -29%, 11% -29%, 12% -29%, 13% -29%, 14% -29%, 15% -29%, 16% -29%, 17% -29%, 18% -29%, 19% -29%, 20% -29%, 21% -29%, 22% -29%, 23% -29%, 24% -29%, 25% -29%, 26% -29%, 27% -29%, 28% -29%, 1% -28%, 2% -28%, 3% -28%, 4% -28%, 5% -28%, 6% -28%, 7% -28%, 8% -28%, 9% -28%, 10% -28%, 11% -28%, 12% -28%, 13% -28%, 14% -28%, 15% -28%, 16% -28%, 17% -28%, 18% -28%, 19% -28%, 20% -28%, 21% -28%, 22% -28%, 23% -28%, 24% -28%, 25% -28%, 26% -28%, 27% -28%, 1% -27%, 2% -27%, 3% -27%, 4% -27%, 5% -27%, 6% -27%, 7% -27%, 8% -27%, 9% -28%, 27%, 10% -27%, 13% -27%, 14% -27%, 15% -28%, 16% -28%, 17% -28%, 18% -28%, and 25% -28%, and 2% -28%, 21% -27%, and 3% -27% 16% -27%, 17% -27%, 18% -27%, 19% -27%, 20% -27%, 21% -27%, 22% -27%, 23% -27%, 24% -27%, 25% -27%, 26% -27%, 1% -26%, 2% -26%, 3% -26%, 4% -26%, 5% -26%, 6% -26%, 7% -26%, 8% -26%, 9% -26%, 10% -26%, 11% -26%, 12% -26%, 13% -26%, 14% -26%, 15% -26%, 16% -26%, 17% -26%, 18% -26%, 19% -26%, 20% -26%, 21% -26%, 22% -26%, 23% -26%, 24% -26%, 25% -26%, 1% -25%, 2% -25%, 3% -25%, 4% -25%, 5% -25%, 6% -25%, 7% -25%, 8% -25%, 9% -25%, 10% -25%, 11% -25%, 12% -25%, 13% -25%, 14% -25%, 15% -25%, 16% -25%, 11% -25%, 18% -25%, 19% -25%, 20% -25%, 21% -25%, 22% -25%, 23% -25%, 24% -25%, 1% -24%, 2% -24%, 3% -24%, 4% -24%, 5% -24%, and, 6% -24%, 7% -24%, 8% -24%, 9% -24%, 10% -24%, 11% -24%, 12% -24%, 13% -24%, 14% -24%, 15% -24%, 16% -24%, 17% -24%, 18% -24%, 19% -24%, 20% -24%, 21% -24%, 22% -24%, 23% -24%, 1% -23%, 2% -23%, 3% -23%, 4% -23%, 5% -23%, 6% -23%, 7% -23%, 8% -23%, 9% -23%, 10% -23%, 11% -23%, 12% -23%, 13% -23%, 14% -23%, 15% -23%, and 16% -23%, 17% -23%, 18% -23%, 19% -23%, 20% -23%, 21% -23%, 22% -23%, 1% -22%, 2% -22%, 3% -22%, 4% -22%, 5% -22%, 6% -22%, 7% -22%, 8% -22%, 9% -22%, 10% -22%, 11% -22%, 12% -22%, 13% -22%, 14% -22%, 15% -22%, 16% -22%, 17% -22%, 18% -22%, 19% -22%, 20% -22%, 21% -22%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 11% -21%, 12% -21%, 13% -21%, 14% -21%, 15% -21%, 16% -21%, 17% -21%, 18% -21%, 19% -21%, 20% -21%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 11% -20%, 12% -20%, 13% -20%, 14% -20%, 15% -20%, 16% -20%, 17% -20%, 18% -20%, 19% -20%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 10% -19%, 11% -19%, 12% -19%, 13% -19%, 14% -19%, 15% -19%, 16% -19%, 17% -19%, 18% -1% -18%, 2% -18%, 3% -18%,4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18%, 9% -18%, 10% -18%, 11% -18%, 12% -18%, and, 13% -18%, 14% -18%, 15% -18%, 16% -18%, 17% -18%, 1% -17%, 2% -17%, 3% -17%, 4% -17%, 5% -17%, 6% -17%, 7% -17%, 8% -17%, 9% -17%, 10% -17%, 11% -17%, 12% -17%, 13% -17%, 14% -17%, 15% -17%, 16% -17%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, 5% -16%, 6% -16%, 7% -16%, 8% -16%, 9% -16%, 10% -16%, 11% -16%, 12% -16%, 11% -16%, and 13% -16%, 14% -16%, 15% -16%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 11% -15%, 12% -15%, 13% -15%, 14% -15%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 11% -14%, 12% -14%, 13% -14%, 1% -13%, 2% -13%, 13% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 11% -13%, 12% -13%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 11% -12%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 1% -10%, 1% -10% and 2% -10%, 3% -10%, 4% -10%, 5% -10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -7%, 1% -6%, 2% -6%, 3% -6%, 4% -6%, and, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -5%, 1% -4%, 2% -4%, 3% -4%, 1% -3%, 2% -3% or 1% -2%.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher temperature tolerance than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. Temperature tolerance can be manifested by increased ethanol production, reduced glycerol production, and increased fermentation rates. In a particular embodiment, the yeast strains and derivatives thereof as described herein can tolerate temperatures of about 20 ℃, about 21 ℃, about 22 ℃, about 23 ℃, about 24 ℃, about 25 ℃, about 26 ℃, about 27 ℃, about 28 ℃, about 29 ℃, about 30 ℃, about 31 ℃, about 32 ℃, about 33 ℃, about 34 ℃, about 35 ℃, about 36 ℃, about 37 ℃, about 38 ℃, about 39 ℃, and/or about 40 ℃. The inventors surprisingly found that the yeast strains and derivatives thereof described herein have a statistically significantly higher temperature tolerance than Y1609 under the same fermentation conditions.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher organic acid tolerance at low pH than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher organic acid tolerance at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher organic acid tolerance at a temperature of 32 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In another specific embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher organic acid tolerance at a temperature of 36 ℃ than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. The inventors surprisingly found that the yeast strains and derivatives thereof described herein have a statistically significantly higher tolerance to organic acids than Y1609 under the same fermentation conditions. Examples of organic acids include, but are not limited to, lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, other carboxylic acids, or combinations thereof.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher ethanol yield at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher ethanol yield at a temperature of 32 ℃ and at low pH of the organic acid than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In another specific embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher ethanol yield at a temperature of 36 ℃ and at a low pH of the organic acid than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. The inventors surprisingly found that under the same fermentation conditions, the yeast strains described herein caused a statistically significantly higher ethanol yield at low pH of the organic acid compared to Y1609. The inventors have also surprisingly found that the derivatives described herein generally lead to statistically significantly higher ethanol yields (e.g., Y1919 in fig. 2-5) at low pH of the organic acid compared to Y1609 under the same conditions.
In some embodiments, the yeast strains and derivatives thereof as described herein produce at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% more ethanol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours relative to a typical yeast strain used for fermentation (e.g., yeast (Saccharomyces) strain Y1609). In some embodiments, the yeast strains and derivatives thereof as described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% ethanol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours relative to typical yeast strains used for fermentation (e.g., yeast(s) (Saccharomyces) strain Y1609). In some embodiments, the yeast strain is selected from the group consisting of yeast (Saccharomyces) strain Y1609, the yeast strains and derivatives thereof as described herein produce about 0.1% -21% (i.e., from about 0.1% to about 21%), 0.2% -21%, 0.3% -21%, 0.4% -21%, 0.5% -21%, 0.6% -21%, 0.7% -21%, 0.8% -21%, 0.9% -21%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 0% -21%, 0.6% -21%, 4% -21%, 0.7% -21%, 0% -21% and derivatives thereof after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of an organic acid for 50 hours 10% -21%, 0.2% -20%, 0.3% -20%, 0.4% -20%, 0.5% -20%, 0.6% -20%, 0.7% -20%, 0.8% -20%, 0.9% -20%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 0.2% -19%, 0.3% -19%, 0.4% -19%, 0.5% -19%, 0.6% -19%, 0.7% -19%, 0.8% -19%, 0.9% -19%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 3% -19%, 4% -5% -19%, 3% -19% and, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 10% -19%, 0.2% -18%, 0.3% -18%, 0.4% -18%, 0.5% -18%, 0.6% -18%, 0.7% -18%, 0.8% -18%, 0.9% -18%, 1% -18%, 2% -18%, 3% -18%, 4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18%, 9% -18%, 10% -18%, 0.2% -17%, 0.3% -17%, 0.4% -17%, 0.5% -17%, 0.6% -17%, 0.7% -17%, 0.8% -17%, 0.9% -17%, 1% -17%, 2% -17%, and so on 3% -17%, 4% -17%, 5% -17%, 6% -17%, 7% -17%, 8% -17%, 9% -17%, 10% -17%, 0.2% -16%, 0.3% -16%, 0.4% -16%, 0.5% -16%, 0.6% -16%, 0.7% -16%, 0.8% -16%, 0.9% -16%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, 5% -16%, 6% -16%, 7% -16%, 8% -16%, 9% -16%, 10% -16%, 0.2% -15%, 0.3% -15%, 0.4% -15%, 0.5% -15%, 0.6% -15%, 0.7% -15%, and, 0.8% -15%, 0.9% -15%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 0.1% -14%, 0.2% -14%, 0.3% -14%, 0.4% -14%, 0.5% -14%, 0.6% -14%, 0.7% -14%, 0.8% -14%, 0.9% -14%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 0.1% -13%, 0.2% -14%, 0.9% -14%, 1% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 0.3% -13%, 0.4% -13%, 0.5% -13%, 0.6% -13%, 0.7% -13%, 0.8% -13%, 0.9% -13%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 0.1% -12%, 0.2% -12%, 0.3% -12%, 0.4% -12%, 0.5% -12%, 0.6% -12%, 0.7% -12%, 0.8% -12%, 0.9% -12%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12% and 0.3% -12% of the composition, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 0.1% -11%, 0.2% -11%, 0.3% -11%, 0.4% -11%, 0.5% -11%, 0.6% -11%, 0.7% -11%, 0.8% -11%, 0.9% -11%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 0.1% -0.2%, 0.1% -0.3%, 0.1% -0.4%, 0.1% -0.5%, 0.1% -0.6%, 0.1% -0.7%, 0.1% -0.8%, 0.1% -0.9%, 0.1% -1%, 0.1% -2%, 0.1% -3%, 0.1% -4%, 0.1% -5%, 0.1% -6%, 0.1% -7%, 0.1% -8% or 0.1% -9% ethanol.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly lower glycerol yield at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly lower glycerol yield at a temperature of 32 ℃ and a low pH of the organic acid than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609. In another particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly lower glycerol yield at a temperature of 36 ℃ and a low pH of the organic acid than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609. The inventors surprisingly found that the yeast strains described herein lead to statistically significantly lower glycerol yields at low pH of organic acids compared to Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein lead to statistically significantly lower glycerol yields (e.g., Y1919 in fig. 2-5) at low pH of the organic acid compared to Y1609 under the same conditions.
In some embodiments, the yeast strains and derivatives thereof as described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, or 22% less glycerol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours relative to a typical yeast strain used for fermentation (e.g., yeast (Saccharomyces) strain Y1609). In some embodiments, the yeast strains and derivatives thereof as described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% less glycerol relative to a typical yeast strain used for fermentation (e.g., yeast strain Y1609) at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and after fermentation at a low pH of an organic acid for 50 hours, the yeast strains and derivatives thereof as described herein produce about 1% -30% (i.e., from about 1% to about 30%), 2% -30%, 3% -30%, 4% -30%, 5% -30%, 6% -30%, 7% -30%, 8% -30%, 9% -30%, 10% -30%, 11% -30%, 12% -30%, 13% -30%, 14% -30%, 15% -30%, 16% -30%, 17% -30%, 18% -30%, 19% -30%, 20% -30%, 21% -30%, 22% -30%, 23% -30%, 24% -30%, 25% -30%, 26% -30%, 27% -30%, 28% -30%, 29% -30%, 1% -29%, 25% -30%, 26% -30% at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of organic acids 2% -29%, 3% -29%, 4% -29%, 5% -29%, 6% -29%, 7% -29%, 8% -29%, 9% -29%, 10% -29%, 11% -29%, 12% -29%, 13% -29%, 14% -29%, 15% -29%, 16% -29%, 17% -29%, 18% -29%, 19% -29%, 20% -29%, 21% -29%, 22% -29%, 23% -29%, 24% -29%, 25% -29%, 26% -29%, 27% -29%, 28% -29%, 1% -28%, 2% -28%, 3% -28%, 4% -28%, 5% -28%, 6% -28%, 7% -28%, 8% -28%, 9% -28%, 10% -28%, 11% -28%, 12% -28%, 13% -28%, 14% -28%, 15% -28%, 16% -28%, 17% -28%, 18% -28%, 19% -28%, 20% -28%, 21% -28%, 22% -28%, 23% -28%, 24% -28%, 25% -28%, 26% -28%, 27% -28%, 1% -27%, 2% -27%, 3% -27%, 4% -27%, 6% -27%, 7% -27%, 27% -27%, 10% -28%, 11% -28%, 12% -28%, 13% -28%, 14% -28%, and 22% -28%, and 23% -28%, and 25% -28% 12% -27%, 13% -27%, 14% -27%, 15% -27%, 16% -27%, 17% -27%, 18% -27%, 19% -27%, 20% -27%, 21% -27%, 22% -27%, 23% -27%, 24% -27%, 25% -27%, 26% -27%, 1% -26%, 2% -26, 3% -26%, 4% -26%, 5% -26%, 6% -26%, 7% -26%, 8% -26%, 9% -26%, 10% -26%, 11% -26%, 12% -26%, 13% -26%, 14% -26%, 15% -26%, 16% -26%, 17% -26%, 18% -26%, and 19% -26%, 20% -26%, 21% -26%, 22% -26%, 23% -26%, 24% -26%, 25% -26%, 1% -25%, 2% -25%, 3% -25%, 4% -25%, 5% -25%, 6% -25%, 7% -25%, 8% -25%, 9% -25%, 10% -25%, 11% -25%, 12% -25%, 13% -25%, 14% -25%, 15% -25%, 16% -25%, 17% -25%, 18% -25%, 19% -25%, 20% -25%, 21% -25%, 22% -25%, 23% -25%, 24% -25%, 1% -24%, and, 2% -24%, 3% -24%, 4% -24%, 5% -24%, 6% -24%, 7% -24%, 8% -24%, 9% -24%, 10% -24%, 11% -24%, 12% -24%, 13% -24%, 14% -24%, 15% -24%, 16% -24%, 17% -24%, 18% -24%, 19% -24%, 20% -24%, 21% -24%, 22% -24%, 23% -24%, 1% -23%, 2% -23%, 3% -23%, 4% -23%, 5% -23%, 6% -23%, 7% -23%, 8% -23%, 9% -23%, 10% -23%, 11% -23%, and 12% -23%, 13% -23%, 14% -23%, 15% -23%, 16% -23%, 17% -23%, 18% -23%, 19% -23%, 20% -23%, 21% -23%, 22% -23%, 1% -22%, 2% -22%, 3% -22%, 4% -22%, 5% -22%, 6% -22%, 7% -22%, 8% -22%, 9% -22%, 10% -22%, 11% -22%, 12% -22%, 13% -22%, 14% -22%, 15% -22%, 16% -22%, 17% -22%, 18% -22%, 19% -22%, 20% -22%, 21% -22%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 11% -21%, 12% -21%, 13% -21%, 14% -21%, 15% -21%, 16% -21%, 17% -21%, 18% -21%, 19% -21%, 20% -21%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 11% -20%, 12% -20%, 13% -20%, and 14% -20%, 15% -20%, 16% -20%, 17% -20%, 18% -20%, 19% -20%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 10% -19%, 11% -19%, 12% -19%, 13% -19%, 14% -19%, 15% -19%, 16% -19%, 17% -19%, 18% -19%, 1% -18%, 2% -18%, 3% -18%, 4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18%, 9% -18%, 10% -18%, 11% -18%, 12% -18%, 13% -18%, 14% -18%, 15% -18%, 16% -18%, 17% -18%, 1% -17%, 2% -17%, 3% -17%, 4% -17%, 5% -17%, 6% -17%, 7% -17%, 8% -17%, 9% -17%, 10% -17%, 11% -17%, 12% -17%, 13% -17%, 14% -17%, 15% -17%, 16% -17%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, 5% -16%, 6% -16%, 7% -16%, 8% -16%, and 9% -16%, 10% -16%, 11% -16%, 12% -16%, 13% -16%, 14% -16%, 15% -16%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 11% -15%, 12% -15%, 13% -15%, 14% -15%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 11% -14%, 12% -14%, 13% -14%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 11% -13%, 12% -13%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 11% -12%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 1% -10%, 2% -10%, 3% -10%, 4% -10%, 5% -10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -7%, 1% -6%, 2% -6%, 3% -6%, 4% -6%, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -5%, 1% -4%, 2% -4%, 3% -4%, 1% -3%, 2% -3% or 1% -2% glycerol.
In some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher fermentation rate at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. In a particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher fermentation rate at a temperature of 32 ℃ and at low pH of the organic acid than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609. In another particular embodiment, the yeast strain and derivatives thereof as described herein have a statistically significantly higher fermentation rate at a temperature of 36 ℃ and at low pH of the organic acid than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609. The inventors surprisingly found that under the same fermentation conditions, the yeast strains described herein caused a statistically significantly higher fermentation rate at low pH of the organic acid compared to Y1609. The inventors surprisingly found that the derivatives described herein lead to statistically significantly higher fermentation rates (e.g., Y1919 in fig. 2-5) at low pH of the organic acid compared to Y1609 under the same conditions.
In some embodiments, the yeast strains and derivatives thereof as described herein have a fermentation rate that is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% higher than a typical yeast strain used for fermentation, e.g., yeast(s) strain Y1609, after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 24 hours. In some embodiments, the yeast strains and derivatives thereof as described herein have a fermentation rate up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% or 50% higher than a typical yeast strain used for fermentation (e.g., yeast(s) strain Y1609) after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of an organic acid for 24 hours. In some embodiments of the present invention, in some embodiments, the yeast strains and derivatives thereof as described herein have a fermentation rate after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid for 24 hours that is about 1% -50% (i.e. from about 1% to about 50%), 2% -50%, 3% -50%, 4% -50%, 5% -50%, 6% -50%, 7% -50%, 8% -50%, 9% -50%, 10% -50%, 11% -50%, 12% -50%, 13% -50%, 14% -50%, 15% -50% higher than typical yeast strains used for fermentation, such as yeast (Saccharomyces) strain Y1609. 16% -50%, 17% -50%, 18% -50%, 19% -50%, 20% -50%, 21% -50%, 22% -50%, 23% -50%, 24% -50%, 25% -50%, 26% -50%, 27% -50%, 28% -50%, 2% -49%, 3% -49%, 4% -49%, 5% -49%, 6% -49%, 7% -49%, 8% -49%, 9% -49%, 10% -49%, 11% -49%, 12% -49%, 13% -49%, 14% -49%, 15% -49%, 16% -49%, 17% -49%, 18% -49%, 19% -49%, 20% -49%, 21% -49%, 22% -49%, 23% -49%, and, 24% -49%, 25% -49%, 26% -49%, 27% -49%, 28% -49%, 2% -48%, 3% -48%, 4% -48%, 5% -48%, 6% -48%, 7% -48%, 8% -48%, 9% -48%, 10% -48%, 11% -48%, 12% -48%, 13% -48%, 14% -48%, 15% -48%, 16% -48%, 17% -48%, 18% -48%, 19% -48%, 20% -48%, 21% -48%, 22% -48%, 23% -48%, 24% -48%, 25% -48%, 26% -48%, 27% -48%, 28% -48%, 2% -47%, and 3% -47%, 4% -47%, 5% -47%, 6% -47%, 7% -47%, 8% -47%, 9% -47%, 10% -47%, 11% -47%, 12% -47%, 13% -47%, 14% -47%, 15% -47%, 16% -47%, 17% -47%, 18% -47%, 19% -47%, 20% -47%, 21% -47%, 22% -47%, 23% -47%, 24% -47%, 25% -47%, 26% -47%, 27% -47%, 28% -47%, 2% -46%, 3% -46%, 4% -46%, 5% -46%, 6% -46%, 7% -46%, 8% -46%, 9% -46%, 10% -46%, 11% -46%, 12% -46%, 13% -46%, 14% -46%, 15% -46%, 16% -46%, 17% -46%, 18% -46%, 19% -46%, 20% -46%, 21% -46%, 22% -46%, 23% -46%, 24% -46%, 25% -46%, 26% -46%, 27% -46%, 28% -46%, 2% -45%, 3% -45%, 4% -45%, 5% -45%, 6% -45%, 7% -45%, 8% -45%, 9% -45%, 10% -45%, 11% -45%, 12% -45%, 13% -45%, and 14% -45%, 15% -45%, 16% -45%, 17% -45%, 18% -45%, 19% -45%, 20% -45%, 21% -45%, 22% -45%, 23% -45%, 24% -45%, 25% -45%, 26% -45%, 27% -45%, 28% -45%, 2% -44%, 3% -44%, 4% -44%, 5% -44%, 6% -44%, 7% -44%, 8% -44%, 9% -44%, 10% -44%, 11% -44%, 12% -44%, 13% -44%, 14% -44%, 15% -44%, 16% -44%, 17% -44%, 18% -44%, 19% -44%, 20% -44%, 21% -44%, 22% -44%, 23% -44%, 24% -44%, 25% -44%, 26% -44%, 27% -44%, 28% -44%, 2% -43%, 3% -43%, 4% -43%, 5% -43%, 6% -43%, 7% -43%, 8% -43%, 9% -43%, 10% -43%, 11% -43%, 12% -43%, 13% -43%, 14% -43%, 15% -43%, 16% -43%, 17% -43%, 18% -43%, 19% -43%, 20% -43%, 21% -43%, 22% -43%, 23% -43%, 24% -43%, 25% -43%, 26% -43%, 27% -43%, 28% -43%, 2% -42%, 3% -42%, 4% -42%, 5% -42%, 6% -42%, 7% -42%, 8% -42%, 9% -42%, 10% -42%, 11% -42%, 12% -42%, 13% -42%, 14% -42%, 15% -42%, 16% -42%, 17% -42%, 18% -42%, 19% -42%, 20% -42%, 21% -42%, 22% -42%, 25% -42%, 27% -42%, 2% -42%, and 2% -42% 3% -41%, 4% -41%, 5% -41%, 6% -41%, 7% -41%, 8% -41%, 9% -41%, 10% -41%, 11% -41%, 12% -41%, 13% -41%, 14% -41%, 15% -41%, 16% -41%, 17% -41%, 18% -41%, 19% -41%, 20% -41%, 21% -41%, 22% -41%, 23% -41%, 24% -41%, 25% -41%, 26% -41%, 27% -41%, 28% -41%, 2% -40%, 3% -40%, 4% -40%, 5% -40%, 6% -40%, 7% -40%, 8% -40%, and 9% -40%, 10% -40%, 11% -40%, 12% -40%, 13% -40%, 14% -40%, 15% -40%, 16% -40%, 17% -40%, 18% -40%, 19% -40%, 20% -40%, 21% -40%, 22% -40%, 23% -40%, 24% -40%, 25% -40%, 26% -40%, 27% -40%, 28% -40%, 2% -39%, 3% -39%, 4% -39%, 5% -39%, 6% -39%, 7% -39%, 8% -39%, 9% -39%, 10% -39%, 11% -39%, 12% -39%, 13% -39%, and, 14% -39%, 15% -39%, 16% -39%, 17% -39%, 18% -39%, 19% -39%, 20% -39%, 21% -39%, 22% -39%, 23% -39%, 24% -39%, 25% -39%, 26% -39%, 27% -39%, 28% -39%, 2% -38%, 3% -38%, 4% -38%, 5% -38%, 6% -38%, 7% -38%, 8% -38%, 9% -38%, 10% -38%, 11% -38%, 12% -38%, 13% -38%, 14% -38%, 15% -38%, 16% -38%, 17% -38%, 18% -38%, 19% -38%, and 20% -38%, 21% -38%, 22% -38%, 23% -38%, 24% -38%, 25% -38%, 26% -38%, 27% -38%, 28% -38%, 2% -37%, 3% -37%, 4% -37%, 5% -37%, 6% -37%, 7% -37%, 8% -37%, 9% -37%, 10% -37%, 11% -37%, 12% -37%, 13% -37%, 14% -37%, 15% -37%, 16% -37%, 17% -37%, 18% -37%, 19% -37%, 20% -37%, 21% -37%, 22% -37%, 23% -37%, 24% -37%, 25% -37%, 26% -37%, 27% -37%, 28% -37%, 2% -36%, 3% -36%, 4% -36%, 5% -36%, 6% -36%, 7% -36%, 8% -36%, 9% -36%, 10% -36%, 11% -36%, 12% -36%, 13% -36%, 14% -36%, 15% -36%, 16% -36%, 17% -36%, 18% -36%, 19% -36%, 20% -36%, 21% -36%, 22% -36%, 23% -36%, 24% -36%, 25% -36%, 26% -36%, 27% -36%, 28% -36%, 2% -35%, 3% -35%, and 4% -35%, 5% -35%, 6% -35%, 7% -35%, 8% -35%, 9% -35%, 10% -35%, 11% -35%, 12% -35%, 13% -35%, 14% -35%, 15% -35%, 16% -35%, 17% -35%, 18% -35%, 19% -35%, 20% -35%, 21% -35%, 22% -35%, 23% -35%, 24% -35%, 25% -35%, 26% -35%, 27% -35%, 28% -35%, 2% -34%, 3% -34%, 4% -34%, 5% -34%, 6% -34%, 7% -34%, 8% -34%, 9% -34%, 10% -34%, 11% -34%, 12% -34%, 13% -34%, 14% -34%, 15% -34%, 16% -34%, 17% -34%, 18% -34%, 19% -34%, 20% -34%, 21% -34%, 22% -34%, 23% -34%, 24% -34%, 25% -34%, 26% -34%, 27% -34%, 28% -34%, 2% -33%, 3% -33%, 4% -33%, 5% -33%, 6% -33%, 7% -33%, 8% -33%, 9% -33%, 10% -33%, 11% -33%, 12% -33%, 13% -33%, 14% -33%, and 15% -33%, 16% -33%, 17% -33%, 18% -33%, 19% -33%, 20% -33%, 21% -33%, 22% -33%, 23% -33%, 24% -33%, 25% -33%, 26% -33%, 27% -33%, 28% -33%, 2% -32%, 3% -32%, 4% -32%, 5% -32%, 6% -32%, 7% -32%, 8% -32%, 9% -32%, 10% -32%, 11% -32%, 12% -32%, 13% -32%, 14% -32%, 15% -32%, 16% -32%, 17% -32%, 18% -32%, 19% -32%, 3% -32%, 4% -32%, 5% -32%, 6% -32%, 7% -32%, 9% -32%, 10% -32%, 11% -32%, 12% -32%, 13% -32%, 14% -32%, 15% -32%, 20% -32%, 21% -32%, 22% -32%, 23% -32%, 24% -32%, 25% -32%, 26% -32%, 27% -32%, 28% -32%, 2% -31%, 3% -31%, 4% -31%, 5% -31%, 6% -31%, 7% -31%, 8% -31%, 9% -31%, 10% -31%, 11% -31%, 12% -31%, 13% -31%, 14% -31%, 15% -31%, 16% -31%, 17% -31%, 18% -31%, 19% -31%, 20% -31%, 21% -31%, 22% -31%, 23% -31%, 24% -31%, 25% -31%, and 26% -31%, 27% -31%, 28% -31%, 2% -30%, 3% -30%, 4% -30%, 5% -30%, 6% -30%, 7% -30%, 8% -30%, 9% -30%, 10% -30%, 11% -30%, 12% -30%, 13% -30%, 14% -30%, 15% -30%, 16% -30%, 17% -30%, 18% -30%, 19% -30%, 20% -30%, 21% -30%, 22% -30%, 23% -30%, 24% -30%, 25% -30%, 26% -30%, 27% -30%, 28% -30%, 29% -30%, 1% -29%, and, 2% -29%, 3% -29%, 4% -29%, 6% -29%, 7% -29%, 8% -29%, 9% -29%, 10% -29%, 11% -29%, 12% -29%, 13% -29%, 14% -29%, 15% -29%, 16% -29%, 17% -29%, 18% -29%, 19% -29%, 20% -29%, 21% -29%, 22% -29%, 23% -29%, 24% -29%, 25% -29%, 26% -29%, 27% -29%, 28% -29%, 1% -28%, 2% -28%, 3% -28%, 4% -28%, 5% -28%, 6% -28%, and 7% -28%, 8% -28%, 9% -28%, 10% -28%, 11% -28%, 12% -28%, 13% -28%, 14% -28%, 15% -28%, 16% -28%, 17% -28%, 18% -28%, 19% -28%, 20% -28%, 21% -28%, 22% -28%, 23% -28%, 24% -28%, 25% -28%, 26% -28%, 27% -28%, 1% -27%, 2% -27%, 3% -27%, 4% -27%, 5% -27%, 6% -27%, 7% -27%, 8% -27%, 9% -27%, 10% -27%, 11% -27%, and, 12% -27%, 13% -27%, 14% -27%, 15% -27%, 16% -27%, 17% -27%, 18% -27%, 19% -27%, 20% -27%, 21% -27%, 22% -27%, 23% -27%, 24% -27%, 25% -27%, 26% -27%, 1% -26%, 2% -26%, 3% -26%, 4% -26%, 5% -26%, 6% -26%, 7% -26%, 8% -26%, 9% -26%, 10% -26%, 11% -26%, 12% -26%, 13% -26%, 14% -26%, 15% -26%, 16% -26%, 17% -26%, 18% -26%, 19% -26%, 20% -26%, 21% -26%, 22% -26%, 23% -26%, 24% -26%, 25% -26%, 1% -25%, 2% -25%, 3% -25%, 4% -25%, 5% -25%, 6% -25%, 7% -25%, 8% -25%, 9% -25%, 10% -25%, 11% -25%, 12% -25%, 13% -25%, 14% -25%, 15% -25%, 16% -25%, 18% -25%, 19% -25%, 20% -25%, 21% -25%, 22% -25%, 23% -25%, 24% -25%, 1% -24% >, 2% -24%, 3% -24%, 4% -24%, 5% -24%, 6% -24%, 7% -24%, 8% -24%, 9% -24%, 10% -24%, 11% -24%, 12% -24%, 13% -24%, 14% -24%, 15% -24%, 16% -24%, 17% -24%, 18% -24%, 19% -24%, 20% -24%, 21% -24%, 22% -24%, 23% -24%, 1% -23%, 2% -23%, 3% -23%, 4% -23%, 5% -23%, 6% -23%, 7% -23%, 8% -23%, 9% -23%, 10% -23%, 11% -23%, and 12% -23%, 13% -23%, 14% -23%, 15% -23%, 16% -23%, 17% -23%, 18% -23%, 19% -23%, 20% -23%, 21% -23%, 22% -23%, 1% -22%, 2% -22%, 3% -22%, 4% -22%, 5% -22%, 6% -22%, 7% -22%, 8% -22%, 9% -22%, 10% -22%, 11% -22%, 12% -22%, 13% -22%, 14% -22%, 15% -22%, 16% -22%, 17% -22%, 18% -22%, 19% -22%, 20% -22%, 21% -22%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 11% -21%, 12% -21%, 13% -21%, 14% -21%, 15% -21%, 16% -21%, 17% -21%, 18% -21%, 19% -21%, 20% -21%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 11% -20%, 12% -20%, 13% -20%, 14% -20%, 15% -20%, 16% -20%, 17% -20%, 18% -20%, 19% -20%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 10% -19%, 11% -19%, 12% -19%, 13% -19%, 14% -19%, 15% -19%, 16% -19%, 17% -19%, 18% -18%, 1% -19%, 18% -18%, 3% -18%,4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18%, and, 9% -18%, 10% -18%, 11% -18%, 12% -18%, 13% -18%, 14% -18%, 15% -18%, 16% -18%, 17% -18%, 1% -17%, 2% -17%, 3% -17%, 4% -17%, 5% -17%, 6% -17%, 7% -17%, 8% -17%, 9% -17%, 10% -17%, 11% -17%, 12% -17%, 13% -17%, 14% -17%, 15% -17%, 16% -17%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, 5% -16%, 6% -16%, 7% -16%, 8% -16%, and 9% -16%, 10% -16%, 11% -16%, 12% -16%, 13% -16%, 14% -16%, 15% -16%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 11% -15%, 12% -15%, 13% -15%, 14% -15%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 11% -14%, 12% -14%, 13% -14%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 11% -13%, 12% -13%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 11% -12%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 1% -10%, 2% -10%, 3% -10%, 4% -10%, 5% -10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -7%, 1% -6%, 2% -6%, 3% -6%, 4% -6%, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -5%, 1% -4%, 2% -4%, 3% -4%, 1% -3%, 2% -3% or 1% -2%.
In some embodiments, the yeast strains and derivatives thereof as described herein have one or more of the defined characteristics described herein. In some embodiments, the yeast strains and derivatives thereof as described herein have one or more of the defined features described herein at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid. In some embodiments, the yeast strains and derivatives thereof as described herein have one or more of the defined features described herein at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at normal pH without an organic acid. For example, in some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher ethanol yield, a statistically significantly lower glycerol yield, a statistically significantly higher fermentation rate, a statistically significantly higher temperature tolerance, and a statistically significantly higher organic acid tolerance than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609; in some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher fermentation rate, produce lower levels of glycerol, and have a statistically significantly higher ethanol yield than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609; in some embodiments, the yeast strains and derivatives thereof as described herein have a statistically significantly higher fermentation rate and produce statistically significantly lower levels of glycerol than typical yeast strains used for fermentation, such as yeast(s) (Saccharomyces) strain Y1609.
The yeast strains and derivatives thereof as described herein may be in any viable form, including crushed, dried (including active dried and instant), compressed, milk forms, and the like. In a particular embodiment, the strain of saccharomyces cerevisiae (Saccharomyces cerevisiae) or derivative thereof is a dry yeast, such as active dry yeast. In another particular embodiment, the strain of Saccharomyces cerevisiae (Saccharomyces cerevisiae) or a derivative thereof is a compressed yeast. In another embodiment, the saccharomyces cerevisiae strain or derivative thereof is a dairy yeast.
a. Yeast strains
One embodiment described herein is a designated yeast (Saccharomyces) strain: y1912 (accession number Y-68003 of NRRL patent deposit); y1913 (deposited under NRRL patent deposit number Y-68004); y1914 (accession number Y-68005 of NRRL patent); y1919 (deposited under NRRL patent deposit number Y-68006); y1923 (under NRRL patent deposit number Y-68007); y1927 (deposited under NRRL patent deposit number Y-68008); y1929 (deposited under NRRL patent deposit number Y-68009). These yeast strains are referred to herein as "the yeast strain", "the yeast (Saccharomyces) strain", or the names thereof (i.e. "Y1912" or "H041", "Y1913" or "H136", "Y1914" or "H138", "Y1919" or "H172", "Y1923" or "H237", "Y1927" or "MM2E2A3", "Y1929" or "MM2E3A 1"). The yeast strains (i.e., Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929) are produced from one or more different yeast (Saccharomyces) strains by one or more methods as shown in FIG. 1. In one aspect, the yeast strains described herein comprise one or more defined characteristics, including higher ethanol yield, lower glycerol yield, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used for fermentation, particularly the yeast strain used in product Fali as compared to yeast strain Y1609. Representative samples of yeast strains have been saved in the agricultural research services patent culture collection (NRRL) under the accession numbers described above, northern regional research center, university street 1815, piolyte, illinois, united states.
b. Yeast strain derivatives
Another embodiment described herein is a derivative of a yeast (Saccharomyces) strain selected from the specified yeast (Saccharomyces) strains: y1912 (accession number Y-68003 of NRRL patent deposit); y1913 (deposited under NRRL patent deposit number Y-68004); y1914 (accession number Y-68005 of NRRL patent); y1919 (deposited under NRRL patent deposit number Y-68006); y1923 (under NRRL patent deposit number Y-68007); y1927 (deposited under NRRL patent deposit number Y-68008); y1929 (deposited under NRRL patent deposit number Y-68009). In one aspect, the derivative comprises one or more defined characteristics including higher ethanol yield, lower glycerol yield, higher temperature tolerance, higher organic acid tolerance and higher fermentation rate than other yeast strains and typical yeast strains used for fermentation, in particular the yeast strain used in product Fali compared to yeast strain Y1609. In another aspect, the derivative may be a parent strain and used to produce other derivatives. For example, Y1912 is a derivative of both strains, but is also a parent strain (e.g., Y1912 is mated with a second strain to produce a Y1913 strain).
c. Mutant yeasts and derivatives thereof
Another embodiment described herein is a mutant of a yeast (Saccharomyces) strain selected from the specified yeast (Saccharomyces) strains: y1912 (accession number Y-68003 of NRRL patent deposit); y1913 (deposited under NRRL patent deposit number Y-68004); y1914 (accession number Y-68005 of NRRL patent); y1919 (deposited under NRRL patent deposit number Y-68006); y1923 (under NRRL patent deposit number Y-68007); y1927 (deposited under NRRL patent deposit number Y-68008); y1929 (deposited under NRRL patent deposit number Y-68009) or a derivative thereof. In one embodiment, the yeast (Saccharomyces) strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 may be derived from one or more different yeast (Saccharomyces) strains by the method shown in FIG. 1D, and thus one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 may be mutant yeast strains. In one aspect, the mutant yeast strains and mutant derivatives comprise one or more defined characteristics, including higher ethanol yield, lower glycerol yield, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used for fermentation, particularly the yeast strain used in product Fali as compared to yeast strain Y1609. In another aspect, the mutant yeast strain and mutant derivatives can be parent strains and used to produce other derivatives. FIG. 1D provides an example of mutagenesis. In one embodiment, the mutant yeast strains and mutant derivatives as described herein are derived from the method shown in fig. 1D.
Mutant yeast strains and mutant derivatives can be prepared by contacting any of the yeast strains described herein with a mutagen. The mutagen may be any mutagen known in the art. For example, the mutagen can be Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine 2 (ICR-170), nitrogen mustard, and the like.
d. Evolved yeast and derivatives thereof
Another embodiment described herein is an evolved yeast strain or derivative of a yeast (Saccharomyces) strain selected from the specified yeast (Saccharomyces) strains: y1912 (accession number Y-68003 of NRRL patent deposit); y1913 (deposited under NRRL patent deposit number Y-68004); y1914 (accession number Y-68005 of NRRL patent); y1919 (deposited under NRRL patent deposit number Y-68006); y1923 (under NRRL patent deposit number Y-68007); y1927 (deposited under NRRL patent deposit number Y-68008); y1929 (deposited under NRRL patent deposit number Y-68009). In one embodiment, the yeast (Saccharomyces) strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 may be derived from one or more different yeast (Saccharomyces) strains by the method shown in FIG. 1C, and thus one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 may be an evolved yeast strain. In one aspect, the evolved yeast strains and the evolved derivatives comprise one or more defined features including higher ethanol yield, lower glycerol yield, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used for fermentation, particularly the yeast strains used in product Fali compared to yeast strain Y1609. In another aspect, the evolved yeast strain and the evolved derivative can be a parent strain and used to produce other derivatives. FIG. 1C provides an example of evolution. In one embodiment, the evolved yeast strains and evolved derivatives as described herein are derived from the method shown in fig. 1C.
The evolved yeast strains and the evolved derivatives can be prepared by applying selection pressure to any of the yeast strains described herein. The selection pressure may be negative (decreasing appearance of the trait) or positive (increasing proportion of the trait). The selection pressure may be constant or may be intermittent. The selection pressure may be applied by varying the presence of resources (e.g., starch and sugar) and/or varying environmental conditions (e.g., temperature, presence of organic acids, pH, and fermentation time).
e. Recombinant yeast and derivatives thereof
Another embodiment described herein is a recombinant yeast strain and a recombinant derivative. The recombinant yeast strains and derivatives may be derived from the saccharomyces cerevisiae (Saccharomyces cerevisiae) strains described herein or derivatives thereof. Recombinant yeast strains can include modifications to inhibit gene expression, enhance gene expression, introduce genes, delete genes, or modify gene sequences. One aspect described herein is a method of preparing a recombinant of a yeast strain or derivative thereof. The method may comprise introducing the nucleic acid into a yeast (Saccharomyces) as described herein using recombinant DNA techniques. Methods for introducing nucleic acids into yeast (Saccharomyces) cells (and in particular yeast (Saccharomyces) strains) are known in the art and are described, for example, in Ausubel et al, (1997), current Protocols in Molecular Biology,2:13.7.1-13.7.7 and Yang and Blhener, (2020), curr Opin Biotechnol.,66:255-266, both of which are incorporated herein by reference. The method may comprise altering the nucleic acid sequence of a yeast (Saccharomyces) or derivative described herein using gene editing or similar techniques.
3. Composition and method for producing the same
Further provided herein are compositions comprising the yeast strains described above or derivatives thereof. In one embodiment, the composition may comprise at least one of the yeast strains described herein, derivatives described herein, or combinations thereof, and naturally occurring and/or non-naturally occurring components. For example, the composition may comprise one or more components selected from surfactants, emulsifiers, gums, swelling agents, antioxidants, starches, metabolites and other processing aids. In one embodiment, the composition may comprise dry yeast of any yeast strain and/or derivative thereof, starch, and an emulsifier. In other embodiments, the composition may comprise milk yeast, glycerol, and xanthan gum of any yeast strain and/or derivative thereof. In one embodiment, an enriched culture of any of the yeast strains described herein is provided, wherein the enriched purity is 90% -99%. In other embodiments, a pure culture of any of the yeast strains described herein is provided, wherein the purity is 100% pure, and therefore no additional yeast is present.
The composition may comprise a yeast (Saccharomyces) as described herein and any suitable surfactant. In one embodiment, the surfactant is an anionic surfactant, a cationic surfactant, and/or a nonionic surfactant.
The composition may comprise yeast (Saccharomyces) as described herein and any suitable emulsifier. In one embodiment, the emulsifier is a fatty acid ester of sorbitan. In one embodiment, the emulsifier is selected from the group consisting of Sorbitol Monostearate (SMS), citric acid esters of mono-diglycerides, polyglycerol esters and fatty acid esters of propylene glycol.
The composition may comprise yeast (Saccharomyces) as described herein and Olindronal SMS, olindronal SK, or Olindronal SPL including the compositions referred to in European patent No. 1,724,336. These products are commercially available from Busaidi, austria for active dry yeasts.
The composition may comprise yeast (Saccharomyces) as described herein and any suitable gum. In one embodiment, the gum is acacia, particularly for milk, compressed and dried yeasts.
The composition may comprise yeast (Saccharomyces) as described herein and any suitable swelling agent. In one embodiment, the swelling agent is methylcellulose or carboxymethylcellulose.
The composition may comprise yeast (Saccharomyces) as described herein and any suitable antioxidant. In one embodiment, the antioxidant is Butyl Hydroxy Anisole (BHA) and/or Butyl Hydroxy Toluene (BHT) or ascorbic acid (vitamin C), particularly for active dry yeasts.
The composition may comprise yeast (Saccharomyces) as described herein and any suitable starch. In one embodiment, the starch is potato starch, corn starch, or pea starch.
The composition may comprise a yeast (Saccharomyces) as described herein and any suitable yeast protectant. In one embodiment, the protective agent is glycerol.
a. Characteristics of the composition
In one aspect, the composition comprises one or more defined characteristics including higher ethanol yield, lower glycerol yield, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast products and typical yeast products used for fermentation, particularly as compared to yeast product Fali. In addition, the compositions described herein can be readily distinguished from other yeast products used in the ethanol industry that do not have the ethanol production capabilities and defined characteristics of the compositions described herein.
In some embodiments, the compositions as described herein are specific for typical yeast products used for fermentation (e.g., fali, ethanolAngel Super/>) Has higher ethanol yield. In some embodiments, the compositions as described herein are at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) compared to typical yeast products used for fermentation (e.g., fali, ethanol- >Angel Super/>) Has higher ethanol yield. In a particular embodiment, the composition as described herein is more effective at a temperature of 32 ℃ than typical yeast products used for fermentation (e.g., fali, ethanoll->Angel Super/>) Has higher ethanol yield. In a further specific embodiment of the present invention, typical yeast products used for fermentation at a temperature ratio of 36℃for compositions as described herein (e.g., fali, ethanol +.>Angel Super/>) Has higher ethanol yield.
In some embodiments, the yeast is a yeast product that is more efficient than typical yeast products used for fermentation (e.g., fali, ethanol Angel Super/>) The compositions as described herein produce up to 50 hours after fermentationAbout 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% less ethanol. In some embodiments, the yeast is used in a fermentation process (e.g., fali, ethanol +.>Angel Super) The compositions as described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% ethanol after 50 hours of fermentation. In some embodiments, the yeast is fermented with respect to a typical yeast product used for fermentation (e.g., fali, ethanol + >Angel Super) Compositions as described herein produce about 0.1% -21% (i.e., from about 0.1% -21%), 0.2% -21%, 0.3% -21%, 0.4% -21%, 0.5% -21%, 0.6% -21%, 0.7% -21%, 0.8% -21%, 0.9% -21%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 0.2% -20%, 0.3% -20%, 0.4% -20%, 0.5% -20%, 0.6% -20%, 0.7% -20%, 0.8% -20%, 0.9% -20%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 0.2% -19%, 0.3% -19%, 0.4% -19%, 0.3% -20%, 0.5% -20%, 0.6% -20%, 0.4% -19%, 0.6% -20%, 0.9% -20%, 0.5% -19%, 0.7% -19% and 0.5% -20% after 50 hours of fermentation％-19％、2％-19％、3％-19％、4％-19％、5％-19％、6％-19％、7％-19％、8％-19％、9％-19％、10％-19％、0.2％-18％、0.3％-18％、0.4％-18％、0.5％-18％、0.6％-18％、0.7％-18％、0.8％-18％、0.9％-18％、1％-18％、2％-18％、3％-18％、4％-18％、5％-18％、6％-18％、7％-18％、8％-18％、9％-18％、10％-18％、0.2％-17％、0.3％-17％、0.4％-17％、0.5％-17％、0.6％-17％、0.7％-17％、0.8％-17％、0.9％-17％、1％-17％、2％-17％、3％-17％、4％-17％、5％-17％、6％-17％、7％-17％、8％-17％、9％-17％、10％-17％、0.2％-16％、0.3％-16％、0.4％-16％、0.5％-16％、0.6％-16％、0.7％-16％、0.8％-16％、0.9％-16％、1％-16％、2％-16％、3％-16％、4％-16％、5％-16％、6％-16％、7％-16％、8％-16％、9％-16％、10％-16％、0.2％-15％、0.3％-15％、0.4％-15％、0.5％-15％、0.6％-15％、0.7％-15％、0.8％-15％、0.9％-15％、1％-15％、2％-15％、3％-15％、4％-15％、5％-15％、6％-15％、7％-15％、8％-15％、9％-15％、10％-15％、0.1％-14％、0.2％-14％、0.3％-14％、0.4％-14％、0.5％-14％、0.6％-14％、0.7％-14％、0.8％-14％、0.9％-14％、1％-14％、2％-14％、3％-14％、4％-14％、5％-14％、6％-14％、7％-14％、8％-14％、9％-14％、10％-14％、0.1％-13％、0.2％-13％、0.3％-13％、0.4％-13％、0.5％-13％、0.6％-13％、0.7％-13％、0.8％-13％、0.9％-13％、1％-13％、2％-13％、3％-13％、4％-13％、5％-13％、6％-13％、7％-13％、8％-13％、9％-13％、10％-13％、0.1％-12％、0.2％-12％、0.3％-12％、0.4％-12％、0.5％-12％、0.6％-12％、0.7％-12％、0.8％-12％、0.9％-12％、1％-12％、2％-12％、3％-12％、4％-12％、5％-12％、6％-12％、7％-12％、8％-12％、9％-12％、10％-12％、0.1％-11％、0.2％-11％、0.3％-11％、0.4％-11％、0.5％-11％、0.6％-11％、0.7％-11％、0.8％-11％0.9% -11%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 0.1% -0.2%, 0.1% -0.3%, 0.1% -0.4%, 0.1% -0.5%, 0.1% -0.6%, 0.1% -0.7%, 0.1% -0.8%, 0.1% -0.9%, 0.1% -1%, 0.1% -2%, 0.1% -3%, 0.1% -4%, 0.1% -5%, 0.1% -6%, 0.1% -7%, 0.1% -8% or 0.1% -9% of ethanol.
In some embodiments, the compositions as described herein are specific for typical yeast products used for fermentation (e.g., fali, ethanolAngel Super/>) With lower glycerol yields. In some embodiments, the compositions as described herein are at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) compared to typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) With lower glycerol yields. In a particular embodiment, the composition as described herein is more effective at a temperature of 32 ℃ than typical yeast products used for fermentation (e.g., fali, ethanoll->Angel Super/>) With lower glycerol yields. In a further specific embodiment of the present invention, typical yeast products used for fermentation at a temperature ratio of 36℃for compositions as described herein (e.g., fali, ethanol +.>Angel Super/>) With lower glycerol yields.
In some embodiments, the yeast is a yeast product that is more efficient than typical yeast products used for fermentation (e.g., fali, ethanol Angel Super/>) The compositions as described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21% or 22% less glycerol after 50 hours of fermentation. In some embodiments, the yeast is fermented with respect to a typical yeast product used for fermentation (e.g., fali, ethanol + >Angel Super/>) The compositions as described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% less glycerol after 50 hours of fermentation. In some embodiments, the yeast is fermented with respect to a typical yeast product used for fermentation (e.g., fali, ethanol +> Angel Super/>) The yeast strains and derivatives thereof as described herein produce about 1% -30% (i.e., from about 1% to about 30%), 2% -30%, 3% -30%, 4% -30%, 5% -30%, 6% -30%, 7% -30%, 8% -30%, 9% -30%, 10% -30%, 11% -30%, 12% -30%, 13% -30%, 14% -30%, 15% -30%, 16% -30%, 17% -30%, 18% -30%, 19% -30%, 20% -30%, 21% -30%, 22% -30%, 23% -30%, 24% -30%, 25% -30%, 26% -30%, and 27% -30%, 28% -30%, 29% -30%, 1% -29%, 2% -29%, 3% -29%, 4% -29%, 5% -29%, 6% -29%, 7% -29%, 8% -29%, 9% -29%, 10% -29%, 11% -29%, 12% -29%, 13% -29%, 14% -29%, 15% -29%, 16% -29%, 17% -29%, 18% -29%, 19% -29%, 20% -29%, 21% -29%, 22% -29%, 23% -29%, 24% -29%, 25% -29%, 26% -29%, 27% -29%, 28% -29%, 1% -28%, 2% -28%, and, 3% -28%, 4% -28%, 5% -28%, 6% -28%, 7% -28%, 8% -28%, 9% -28%, 10% -28%, 11% -28%, 12% -28%, 13% -28%, 14% -28%, 15% -28%, 16% -28%, 17% -28%, 18% -28%, 19% -28%, 20% -28%, 21% -28%, 22% -28%, 23% -28%, 24% -28%, 25% -28%, 26% -28%, 27% -28%, 1% -27%, 2% -27%, 3% -27%, 4% -27%, 5% -27%, 6% -27%, 7% -27%, 8% -27%, 9% -27%, 10% -27%, 11% -27%, 12% -27%, 13% -27%, 14% -27%, 15% -27%, 16% -27%, 17% -27%, 18% -27%, 19% -27%, 20% -27%, 21% -27%, 22% -27%, 23% -27%, 24% -27%, 25% -27%, 26% -27%, 1% -26%, 2% -26%, 3% -26%, 4% -26%, 5% -26%, 6% -26%, 7% -26%, 9% -26%, 10% -26%, 11% -27%, 12% -14% -27%, 13% -27%, 14% -27%, 15% -27%, 16% -27%, and 25% -25%, and 25% -27% 15% -26%, 16% -26%, 17% -26%, 18% -26%, 19% -26%, 20% -26%, 21% -26%, 22% -26%, 23% -26%, 24% -26%, 25% -26%, and, 1％-25％、2％-25％、3％-25％、4％-25％、5％-25％、6％-25％、7％-25％、8％-25％、9％-25％、10％-25％、11％-25％、12％-25％、13％-25％、14％-25％、15％-25％、16％-25％、17％-25％、18％-25％、19％-25％、20％-25％、21％-25％、22％-25％、23％-25％、24％-25％、1％-24％、2％-24％、3％-24％、4％-24％、5％-24％、6％-24％、7％-24％、8％-24％、9％-24％、10％-24％、11％-24％、12％-24％、13％-24％、14％-24％、15％-24％、16％-24％、17％-24％、18％-24％、19％-24％、20％-24％、21％-24％、22％-24％、23％-24％、1％-23％、2％-23％、3％-23％、4％-23％、5％-23％、6％-23％、7％-23％、8％-23％、9％-23％、10％-23％、11％-23％、12％-23％、13％-23％、14％-23％、15％-23％、16％-23％、17％-23％、18％-23％、19％-23％、20％-23％、21％-23％、22％-23％、1％-22％、2％-22％、3％-22％、4％-22％、5％-22％、6％-22％、7％-22％、8％-22％、9％-22％、10％-22％、11％-22％、12％-22％、13％-22％、14％-22％、15％-22％、16％-22％、17％-22％、18％-22％、19％-22％、20％-22％、21％-22％、1％-21％、2％-21％、3％-21％、4％-21％、5％-21％、6％-21％、7％-21％、8％-21％、9％-21％、10％-21％、11％-21％、12％-21％、13％-21％、14％-21％、15％-21％、16％-21％、17％-21％、18％-21％、19％-21％、20％-21％、1％-20％、2％-20％、3％-20％、4％-20％、5％-20％、6％-20％、7％-20％、8％-20％、9％-20％、10％-20％、11％-20％、12％-20％、13％-20％、14％-20％、15％-20％、16％-20％、17％-20％、18％-20％、19％-20％、1％-19％、2％-19％、3％-19％、4％-19％、5％-19％、6％-19％、7％-19％、8％-19％、9％-19％、10％-19％、11％-19％、12％-19％、13％-19％、14％-19％、15％-19％、16％-19％、17％-19％、18％-19％、1％-18％、2％-18％、3％-18％、4％-18％、5-18%, 6% -18%, 7% -18%, 8% -18%, 9% -18%, 10% -18%, 11% -18%, 12% -18%, 13% -18%, 14% -18%, 15% -18%, 16% -18%, 17% -18%, 1% -17%, 2% -17%, 3% -17%, 4% -17%, 5% -17%, 6% -17%, 7% -17%, 8% -17%, 9% -17%, 10% -17%, 11% -17%, 12% -17%, 13% -17%, 14% -17%, 15% -17%, 16% -17%, 1% -16%, 2% -16%, 3% -16%, 4% -16%, and 5% -16%, 6% -16%, 7% -16%, 8% -16%, 9% -16%, 10% -16%, 11% -16%, 12% -16%, 13% -16%, 14% -16%, 15% -16%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 11% -15%, 12% -15%, 13% -15%, 14% -15%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 11% -14%, 12% -14%, 13% -14%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 11% -13%, 12% -13%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 11% -12%, 1% -11%, 2% -11%, 3% -11%, 11% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 1% -10%, 2% -10%, 3% -10%, 4% -10%, 5% -10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 7% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -7%, 1% -6%, 2% -6%, 3% -6%, 4% -6%, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -5%, 1% -4%, 2% -4%, 3% -4%, 1% -3%, 2% -3% or 1% -2% glycerol.
In some embodiments, the compositions as described herein are used in fermentationIs typical of yeast products (e.g., fali, ethanolAngel Super/>) Has higher fermentation rate. In some embodiments, the compositions as described herein are at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) compared to typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) Has higher fermentation rate. In a particular embodiment, the composition as described herein is more effective at a temperature of 32 ℃ than typical yeast products used for fermentation (e.g., fali, ethanoll->Angel Super/>) Has higher fermentation rate. In a further specific embodiment of the present invention, typical yeast products used for fermentation at a temperature ratio of 36℃for compositions as described herein (e.g., fali, ethanol +.>Angel Super/>) Has higher fermentation rate.
In some embodiments, the compositions as described herein have a fermentation rate after 24 hours of fermentation that is greater than that of a typical yeast product used for fermentation (e.g., fali, ethanolAngel Super/>) At least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% higher. In some embodiments, the compositions as described herein have a fermentation rate after 24 hours of fermentation that is greater than that of a typical yeast product used for fermentation (e.g., fali, ethanol + - >Angel Super/>) Up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% or 50% higher. In some embodiments, the compositions as described herein have a fermentation rate after 24 hours of fermentation that is greater than that of a typical yeast product used for fermentation (e.g., fali, ethanol + ->Angel Super/>) From about 1% -50% (i.e., from about 1% -50%), 2% -50%, 3% -50%, 4% -50%, 5% -50%, 6% -50%, 7% -50%, 8% -50%, 9% -50%, 10% -50%, 11% -50%, 12% -50%, 13% -50%, 14% -50%, 15% -50%, 16% -50%, 17% -50%, 18% -50%, 19% -50%, 20% -50%, 21% -50%, 22% -50%, 23% -50%, 24% -50%, 25% -50%, 26% -50%, 27% -50%, 28% -50%, 2% -49%, 3% -49%, 4% -49%, 5% -49%, 6%-49％、7％-49％、8％-49％、9％-49％、10％-49％、11％-49％、12％-49％、13％-49％、14％-49％、15％-49％、16％-49％、17％-49％、18％-49％、19％-49％、20％-49％、21％-49％、22％-49％、23％-49％、24％-49％、25％-49％、26％-49％、27％-49％、28％-49％、2％-48％、3％-48％、4％-48％、5％-48％、6％-48％、7％-48％、8％-48％、9％-48％、10％-48％、11％-48％、12％-48％、13％-48％、14％-48％、15％-48％、16％-48％、17％-48％、18％-48％、19％-48％、20％-48％、21％-48％、22％-48％、23％-48％、24％-48％、25％-48％、26％-48％、27％-48％、28％-48％、2％-47％、3％-47％、4％-47％、5％-47％、6％-47％、7％-47％、8％-47％、9％-47％、10％-47％、11％-47％、12％-47％、13％-47％、14％-47％、15％-47％、16％-47％、17％-47％、18％-47％、19％-47％、20％-47％、21％-47％、22％-47％、23％-47％、24％-47％、25％-47％、26％-47％、27％-47％、28％-47％、2％-46％、3％-46％、4％-46％、5％-46％、6％-46％、7％-46％、8％-46％、9％-46％、10％-46％、11％-46％、12％-46％、13％-46％、14％-46％、15％-46％、16％-46％、17％-46％、18％-46％、19％-46％、20％-46％、21％-46％、22％-46％、23％-46％、24％-46％、25％-46％、26％-46％、27％-46％、28％-46％、2％-45％、3％-45％、4％-45％、5％-45％、6％-45％、7％-45％、8％-45％、9％-45％、10％-45％、11％-45％、12％-45％、13％-45％、14％-45％、15％-45％、16％-45％、17％-45％、18％-45％、19％-45％、20％-45％、21％-45％、22％-45％、23％-45％、24％-45％、25％-45％、26％-45％、27％-45％、28％-45％、2％-44％、3％-44％、4％-44％、5％-44％、6％-44％、7％-44％、8％-44％、9％-44％、10％-44％、11％-44％、12％-44％、13％-44％、14％-44％、15％-44％、16％-44％、17％-44％、18％-44％、19％-44％、20％-44％、21％-44％、22％-44％、23％-44％、24％-44％、25％-44％、26％-44％、27％-44％、28％-44％、2％-43％、3％-43％、4％-43％、5％-43％、6％-43％、7％-43％、8％-43％、9％-43％、10％-43％、11％-43％、12％-43％、13％-43％、14％-43％、15％-43％、16％-43％、17％-43％、18％-43％、19％-43％、20％-43％、21％-43％、22％-43％、23％-43％、24％-43％、25％-43％、26％-43％、27％-43％、28％-43％、2％-42％、3％-42％、4％-42％、5％-42％、6％-42％、7％-42％、8％-42％、9％-42％、10％-42％、11％-42％、12％-42％、13％-42％、14％-42％、15％-42％、16％-42％、17％-42％、18％-42％、19％-42％、20％-42％、21％-42％、22％-42％、23％-42％、24％-42％、25％-42％、26％-42％、27％-42％、28％-42％、2％-41％、3％-41％、4％-41％、5％-41％、6％-41％、7％-41％、8％-41％、9％-41％、10％-41％、11％-41％、12％-41％、13％-41％、14％-41％、15％-41％、16％-41％、17％-41％、18％-41％、19％-41％、20％-41％、21％-41％、22％-41％、23％-41％、24％-41％、25％-41％、26％-41％、27％-41％、28％-41％、2％-40％、3％-40％、4％-40％、5％-40％、6％-40％、7％-40％、8％-40％、9％-40％、10％-40％、11％-40％、12％-40％、13％-40％、14％-40％、15％-40％、16％-40％、17％-40％、18％-40％、19％-40％、20％-40％、21％-40％、22％-40％、23％-40％、24％-40％、25％-40％、26％-40％、27％-40％、28％-40％、2％-39％、3％-39％、4％-39％、5％-39％、6％-39％、7％-39％、8％-39％、9％-39％、10％-39％、11％-39％、12％-39％、13％-39％、14％-39％、15％-39％、16％-39％、17％-39％、18％-39％、19％-39％、20％-39％、21％-39％、22％-39％、23％-39％、24％-39％、25％-39％、26％-39％、27％-39％、28％-39％、2％-38％、3％-38％、4％-38％、5％-38％、6％-38％、7％-38％、8％-38％、9％-38％、10％-38％、11％-38％、12％-38％、13％-38％、14％-38％、15％-38％、16％-38％、17％-38％、18％-38％、19％-38％、20％-38％、21％-38％、22％-38％、23％-38％、24％-38％、25％-38％、26％-38％、27％-38％、28％-38％、2％-37％、3％-37％、4％-37％、5％-37％、6％-37％、7％-37％、8％-37％、9％-37％、10％-37％、11％-37％、12％-37％、13％-37％、14％-37％、15％-37％、16％-37％、17％-37％、18％-37％、19％-37％、20％-37％、21％-37％、22％-37％、23％-37％、24％-37％、25％-37％、26％-37％、27％-37％、28％-37％、2％-36％、3％-36％、4％-36％、5％-36％、6％-36％、7％-36％、8％-36％、9％-36％、10％-36％、11％-36％、12％-36％、13％-36％、14％-36％、15％-36％、16％-36％、17％-36％、18％-36％、19％-36％、20％-36％、21％-36％、22％-36％、23％-36％、24％-36％、25％-36％、26％-36％、27％-36％、28％-36％、2％-35％、3％-35％、4％-35％、5％-35％、6％-35％、7％-35％、8％-35％、9％-35％、10％-35％、11％-35％、12％-35％、13％-35％、14％-35％、15％-35％、16％-35％、17％-35％、18％-35％、19％-35％、20％-35％、21％-35％、22％-35％、23％-35％、24％-35％、25％-35％、26％-35％、27％-35％、28％-35％、2％-34％、3％-34％、4％-34％、5％-34％、6％-34％、7％-34％、8％-34％、9％-34％、10％-34％、11％-34％、12％-34％、13％-34％、14％-34％、15％-34％、16％-34％、17％-34％、18％-34％、19％-34％、20％-34％、21％-34％、22％-34％、23％-34％、24％-34％、25％-34％、26％-34％、27％-34％、28％-34％、2％-33％、3％-33％、4％-33％、5％-33％、6％-33％、7％-33％、8％-33％、9％-33％、10％-33％、11％-33％、12％-33％、13％-33％、14％-33％、15％-33％、16％-33％、17％-33％、18％-33％、19％-33％、20％-33％、21％-33％、22％-33％、23％-33％、24％-33％、25％-33％、26％-33％、27％-33％、28％-33％、2％-32％、3％-32％、4％-32％、5％-32％、6％-32％、7％-32％、8％-32％、9％-32％、10％-32％、11％-32％、12％-32％、13％-32％、14％-32％、15％-32％、16％-32％、17％-32％、18％-32％、19％-32％、20％-32％、21％-32％、22％-32％、23％-32％、24％-32％、25％-32％、26％-32％、27％-32％、28％-32％、2％-31％、3％-31％、4％-31％、5％-31％、6％-31％、7％-31％、8％-31％、9％-31％、10％-31％、11％-31％、12％-31％、13％-31％、14％-31％、15％-31％、16％-31％、17％-31％、18％-31％、19％-31％、20％-31％、21％-31％、22％-31％、23％-31％、24％-31％、25％-31％、26％-31％、27％-31％、28％-31％、2％-30％、3％-30％、4％-30％、5％-30％、6％-30％、7％-30％、8％-30％、9％-30％、10％-30％、11％-30％、12％-30％、13％-30％、14％-30％、15％-30％、16％-30％、17％-30％、18％-30％、19％-30％、20％-30％、21％-30％、22％-30％、23％-30％、24％-30％、25％-30％、26％-30％、27％-30％、28％-30％、29％-30％、1％-29％、2％-29％、3％-29％、4％-29％、％-29％、6％-29％、7％-29％、8％-29％、9％-29％、10％-29％、11％-29％、12％-29％、13％-29％、14％-29％、15％-29％、16％-29％、17％-29％、18％-29％、19％-29％、20％-29％、21％-29％、22％-29％、23％-29％、24％-29％、25％-29％、26％-29％、27％-29％、28％-29％、1％-28％、2％-28％、3％-28％、4％-28％、5％-28％、6％-28％、7％-28％、8％-28％、9％-28％、10％-28％、11％-28％、12％-28％、13％-28％、14％-28％、15％-28％、16％-28％、17％-28％、18％-28％、19％-28％、20％-28％、21％-28％、22％-28％、23％-28％、24％-28％、25％-28％、26％-28％、27％-28％、1％-27％、2％-27％、3％-27％、4％-27％、5％-27％、6％-27％、7％-27％、8％-27％、9％-27％、10％-27％、11％-27％、12％-27％、13％-27％、14％-27％、15％-27％、16％-27％、17％-27％、18％-27％、19％-27％、20％-27％、21％-27％、22％-27％、23％-27％、24％-27％、25％-27％、26％-27％、1％-26％、2％-26％、3％-26％、4％-26％、5％-26％、6％-26％、7％-26％、8％-26％、9％-26％、10％-26％、11％-26％、12％-26％、13％-26％、14％-26％、15％-26％、16％-26％、17％-26％、18％-26％、19％-26％、20％-26％、21％-26％、22％-26％、23％-26％、24％-26％、25％-26％、1％-25％、2％-25％、3％-25％、4％-25％、5％-25％、6％-25％、7％-25％、8％-25％、9％-25％、10％-25％、11％-25％、12％-25％、13％-25％、14％-25％、15％-25％、16％-25％、％-25％、18％-25％、19％-25％、20％-25％、21％-25％、22％-25％、23％-25％、24％-25％、1％-24％、2％-24％、3％-24％、4％-24％、5％-24％、6％-24％、7％-24％、8％-24％、9％-24％、10％-24％、11％-24％、12％-24％、13％-24％、14％-24％、15％-24％、16％-24％、17％-24％、18％-24％、19％-24％、20％-24％、21％-24％、22％-24％、23％-24％、1％-23％、2％-23％、3％-23％、4％-23％、5％-23％、6％-23％、7％-23％、8％-23％、9％-23％、10％-23％、11％-23％、12％-23％、13％-23％、14％-23％、15％-23％、16％-23％、17％-23％、18％-23％、19％-23％、20％-23％、21％-23％、22％-23％、1％-22％、2％-22％、3％-22％、4％-22％、5％-22％、6％-22％、7％-22％、8％-22％、9％-22％、10％-22％、11％-22％、12％-22％、13％-22％、14％-22％、15％-22％、16％-22％、17％-22％、18％-22％、19％-22％、20％-22％、21％-22％、1％-21％、2％-21％、3％-21％、4％-21％、5％-21％、6％-21％、7％-21％、8％-21％、9％-21％、10％-21％、11％-21％、12％-21％、13％-21％、14％-21％、15％-21％、16％-21％、17％-21％、18％-21％、19％-21％、20％-21％、1％-20％、2％-20％、3％-20％、4％-20％、5％-20％、6％-20％、7％-20％、8％-20％、9％-20％、10％-20％、11％-20％、12％-20％、13％-20％、14％-20％、15％-20％、16％-20％、17％-20％、18％-20％、19％-20％、1％-19％、2％-19％、3％-19％、4％-19％、5％-19％、6％-19％、7％-19％、8％-19％、9％-19％、10％-19％、11％-19％、12％-19％、13％-19％、14％-19％、15％-19％、16％-19％、17％-19％、18％-19％、1％-18％、2％-18％、3％-18％、4％-18％、5％-18％、6％-18％、7％-18％、8％-18％、9％-18％、10％-18％、11％-18％、12％-18％、13％-18％、14％-18％、15％-18％、16％-18％、17％-18％、1％-17％、2％-17％、3％-17％、4％-17％、5％-17％、6％-17％、7％-17％、8％-17％、9％-17％、10％-17％、11％-17％、12％-17％、13％-17％、14％-17％、15％-17％、16％-17％、1％-16％、2％-16％、3％-16％、4％-16％、5％-16％、6％-16％、7％-16％、8％-16％、9％-16％、10％-16％、11％-16％、12％-16％、13％-16％、14％-16％、15％-16％、1％-15％、2％-15％、3％-15％、4％-15％、5％-15％、6％-15％、7％-15％、8％-15％、9％-15％、10％-15％、11％-15％、12％-15％、13％-15％、14％-15％、1％-14％、2％-14％、3％-14％、4％-14％、5％-14％、6％-14％、7％-14％、8％-14％、9％-14％、10％-14％、11％-14％、12％-14％、13％-14％、1％-13％、2％-13％、3％-13％、4％-13％、5％-13％、6％-13％、7％-13％、8％-13％、9％-13％、10％-13％、11％-13％、12％-13％、1％-12％、2％-12％、3％-12％、4％-12％、5％-12％、6％-12％、7％-12％、8％-12％、9％-12％、10％-12％、11％-12％、1％-11％、2％-11％、3％-11％、4％-11％、5％-11％、6％-11％、7％-11％、8％-11％、9％-11％、10％-11％、1％-10％、2％-10％、3％-10％、4％-10％、5％-10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -7%, 1% -6%, 2% -6%, 3% -6%, 4% -6%, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -5%, 1% -4%, 2% -4%, 3% -4%, 1% -3%, 2% -3% or 1% -2%.
In some embodiments, the compositions as described herein are specific for typical yeast products used for fermentation (e.g., fali, ethanolAngel Super/>) Has higher temperature tolerance. Temperature tolerance can be manifested by increased ethanol production, reduced glycerol production, and increased fermentation rates. In a particular embodiment, the composition as described herein can withstand temperatures of about 20 ℃, about 21 ℃, about 22 ℃, about 23 ℃, about 24 ℃, about 25 ℃, about 26 ℃, about 27 ℃, about 28 ℃, about 29 ℃, about 30 ℃, about 31 ℃, about 32 ℃, about 33 ℃, about 34 ℃, about 35 ℃, about 36 ℃, about 37 ℃, about 38 ℃, about 39 ℃, and/or about 40 ℃.
In some embodiments, the compositions as described herein are less stable at low pH than typical yeast products used for fermentation (e.g., fali, ethanolAngel Super/>) Has higher organic acid tolerance. In some embodiments, the compositions as described herein are at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) compared to typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) Has higher organic acid tolerance. In a particular embodiment, the compositions as described herein are more effective at a temperature of 32 ℃ than typical yeast products used for fermentation (e.g., fali, ethanol- >Angel Super/>) Has significantly higher organic acid tolerance. In another particular embodiment, the compositions as described herein are more effective than typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) Has significantly higher organic acid tolerance.
In some embodiments, the compositions as described herein are specific to typical yeast products used for fermentation (e.g., fali, ethanol) at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid Angel Super/>) With significantly higher ethanol yields. In a particular embodiment, the composition as described herein is used for fermentation at a temperature of 32 ℃ and at a low pH of the organic acidIs a typical yeast product of (e.g., fali, ethanol +)>Angel Super/>) With significantly higher ethanol yields. In another particular embodiment, the compositions as described herein are specific for typical yeast products used for fermentation at a temperature of 36 ℃ and at low pH of the organic acid (e.g., fali, ethanol->Angel Super) With significantly higher ethanol yields.
In some embodiments, the yeast is a yeast product that is more efficient than typical yeast products used for fermentation (e.g., fali, ethanol Angel Super/>) The compositions as described herein produce at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% more ethanol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours. In some embodiments, the yeast is fermented with respect to a typical yeast product used for fermentation (e.g., fali, ethanol + >Angel Super/>) The compositions as described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0% or 21.0% ethanol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours. In some embodiments, the yeast is a yeast product that is more efficient than typical yeast products used for fermentation (e.g., fali, ethanolAngel Super/>) The compositions as described herein produce about 0.1% -21% (i.e., from about 0.1% to about 21%), 0.2% -21%, 0.3% -21%, 0.4% -21%, 0.5% -21%, 0.6% -21%, 0.7% -21%, 0.8% -21%, 0.9% -21%, 1% -21%, 2% -21%, 3% -21%, 4% -21%, 5% -21%, 6% -21%, 7% -21%, 8% -21%, 9% -21%, 10% -21%, 0.2% -20%, 0.3% -20%, 0.8% -21%, 0.3% -21% after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of an organic acid for 50 hours 0.4% -20%, 0.5% -20%, 0.6% -20%, 0.7% -20%, 0.8% -20%, 0.9% -20%, 1% -20%, 2% -20%, 3% -20%, 4% -20%, 5% -20%, 6% -20%, 7% -20%, 8% -20%, 9% -20%, 10% -20%, 0.2% -19%, 0.3% -19%, 0.4% -19%, 0.5% -19%, 0.6% -19%, 0.7% -19%, 0.8% -19%, 0.9% -19%, 1% -19%, 2% -19%, 3% -19%, 4% -19%, 5% -19%, 6% -19%, 7% -19%, 8% -19%, 9% -19%, 10% -19%, 0.2% -18%, 0.3% -19%, 0.5% -19%, 0.9% -1% -19%, 2% -19%, 3% -19%, 4% -19%, 7% -19%, 8% -19%, 9% -10% -19%, and 0.2% -18% and the like, 0.3% -18%, 0.4% -18%, 0.5% -18%, 0.6% -18%, 0.7% -18%, 0.8% -18%, 0.9% -18%, 1% -18%, 2% -18%, 3% -18%, 4% -18%, 5% -18%, 6% -18%, 7% -18%, 8% -18 ％、9％-18％、10％-18％、0.2％-17％、0.3％-17％、0.4％-17％、0.5％-17％、0.6％-17％、0.7％-17％、0.8％-17％、0.9％-17％、1％-17％、2％-17％、3％-17％、4％-17％、5％-17％、6％-17％、7％-17％、8％-17％、9％-17％、10％-17％、0.2％-16％、0.3％-16％、0.4％-16％、0.5％-16％、0.6％-16％、0.7％-16％、0.8％-16％、0.9％-16％、1％-16％、2％-16％、3％-16％、4％-16％、5％-16％、6％-16％、7％-16％、8％-16％、9％-16％、10％-16％、0.2％-15％、0.3％-15％、0.4％-15％、0.5％-15％、0.6％-15％、0.7％-15％、0.8％-15％、0.9％-15％、1％-15％、2％-15％、3％-15％、4％-15％、5％-15％、6％-15％、7％-15％、8％-15％、9％-15％、10％-15％、0.1％-14％、0.2％-14％、0.3％-14％、0.4％-14％、0.5％-14％、0.6％-14％、0.7％-14％、0.8％-14％、0.9％-14％、1％-14％、2％-14％、3％-14％、4％-14％、5％-14％、6％-14％、7％-14％、8％-14％、9％-14％、10％-14％、0.1％-13％、0.2％-13％、0.3％-13％、0.4％-13％、0.5％-13％、0.6％-13％、0.7％-13％、0.8％-13％、0.9％-13％、1％-13％、2％-13％、3％-13％、4％-13％、5％-13％、6％-13％、7％-13％、8％-13％、9％-13％、10％-13％、0.1％-12％、0.2％-12％、0.3％-12％、0.4％-12％、0.5％-12％、0.6％-12％、0.7％-12％、0.8％-12％、0.9％-12％、1％-12％、2％-12％、3％-12％、4％-12％、5％-12％、6％-12％、7％-12％、8％-12％、9％-12％、10％-12％、0.1％-11％、0.2％-11％、0.3％-11％、0.4％-11％、0.5％-11％、0.6％-11％、0.7％-11％、0.8％-11％、0.9％-11％、1％-11％、2％-11％、3％-11％、4％-11％、5％-11％、6％-11％、7％-11％、8％-11％、9％-11％、10％-11％、0.1％-0.2％、0.1％-0.3％、0.1％-0.4％、0.1％-0.5％、0.1％-0.6％、0.1％-0.7％、0.1％-0.8％、0.1％-0.9％、0.1％-1％、0.1％-2％、0.1％-3％、0.1％-4％、0.1％-5％、0.1% -6%, 0.1% -7%, 0.1% -8% or 0.1% -9% ethanol.
In some embodiments, the compositions as described herein are specific to typical yeast products used for fermentation (e.g., fali, ethanol) at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid Angel Super/>) With lower glycerol yields. In a particular embodiment, the composition as described herein is specific to typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) With lower glycerol yields. In another particular embodiment, the composition as described herein is specific to typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) With lower glycerol yields.
In some embodiments, the yeast is a yeast product that is more efficient than typical yeast products used for fermentation (e.g., fali, ethanol Angel Super/>) The compositions as described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21% or 22% less glycerol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours. In some embodiments, the yeast is a yeast product that is more efficient than typical yeast products used for fermentation (e.g., fali, ethanol Angel Super/>) The compositions as described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% less glycerol after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid for 50 hours. In some embodiments, the yeast is fermented with respect to a typical yeast product used for fermentation (e.g., fali, ethanol +>Angel Super/>) The compositions as described herein produce about 1% -30% (i.e., from about 1% to about 30%), 2% -30%, 3% -30%, 4% -30%, 5% -30%, 6% -30%, 7% -30%, 8% -30%, 9% -30%, 10% -30%, 11% -30%, 12% -30%, 13% -30%, 14% -30%, 15% -30%, 16% -30%, 17% -30%, 18% -30%, 19% -30%, 20% -30%, 21% -30%, 22% less after fermentation at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of an organic acid for 50 hours-30％、23％-30％、24％-30％、25％-30％、26％-30％、27％-30％、28％-30％、29％-30％、1％-29％、2％-29％、3％-29％、4％-29％、5％-29％、6％-29％、7％-29％、8％-29％、9％-29％、10％-29％、11％-29％、12％-29％、13％-29％、14％-29％、15％-29％、16％-29％、17％-29％、18％-29％、19％-29％、20％-29％、21％-29％、22％-29％、23％-29％、24％-29％、25％-29％、26％-29％、27％-29％、28％-29％、1％-28％、2％-28％、3％-28％、4％-28％、5％-28％、6％-28％、7％-28％、8％-28％、9％-28％、10％-28％、11％-28％、12％-28％、13％-28％、14％-28％、15％-28％、16％-28％、17％-28％、18％-28％、19％-28％、20％-28％、21％-28％、22％-28％、23％-28％、24％-28％、25％-28％、26％-28％、27％-28％、1％-27％、2％-27％、3％-27％、4％-27％、5％-27％、6％-27％、7％-27％、8％-27％、9％-27％、10％-27％、11％-27％、12％-27％、13％-27％、14％-27％、15％-27％、16％-27％、17％-27％、18％-27％、19％-27％、20％-27％、21％-27％、22％-27％、23％-27％、24％-27％、25％-27％、26％-27％、1％-26％、2％-26、3％-26％、4％-26％、5％-26％、6％-26％、7％-26％、8％-26％、9％-26％、10％-26％、11％-26％、12％-26％、13％-26％、14％-26％、15％-26％、16％-26％、17％-26％、18％-26％、19％-26％、20％-26％、21％-26％、22％-26％、23％-26％、24％-26％、25％-26％、1％-25％、2％-25％、3％-25％、4％-25％、5％-25％、6％-25％、7％-25％、8％-25％、9％-25％、10％-25％、11％-25％、12％-25％、13％-25％、14％-25％、15％-25％、16％-25％、17％-25％、18％-25％、19％-25％、20％-25％、21％-25％、22％-25％、23％-25％、24％-25％、1％-24％、2％-24％、3％-24％、4％-24％、5％-24％、6％-24％、7％-24％、8％-24％、9％-24％、10％-24％、11％-24％、12％-24％、13％-24％、14％-24％、15％-24％、16％-24％、17％-24％、18％-24％、19％-24％、20％-24％、21％-24％、22％-24％、23％-24％、1％-23％、2％-23％、3％-23％、4％-23％、5％-23％、6％-23％、7％-23％、8％-23％、9％-23％、10％-23％、11％-23％、12％-23％、13％-23％、14％-23％、15％-23％、16％-23％、17％-23％、18％-23％、19％-23％、20％-23％、21％-23％、22％-23％、1％-22％、2％-22％、3％-22％、4％-22％、5％-22％、6％-22％、7％-22％、8％-22％、9％-22％、10％-22％、11％-22％、12％-22％、13％-22％、14％-22％、15％-22％、16％-22％、17％-22％、18％-22％、19％-22％、20％-22％、21％-22％、1％-21％、2％-21％、3％-21％、4％-21％、5％-21％、6％-21％、7％-21％、8％-21％、9％-21％、10％-21％、11％-21％、12％-21％、13％-21％、14％-21％、15％-21％、16％-21％、17％-21％、18％-21％、19％-21％、20％-21％、1％-20％、2％-20％、3％-20％、4％-20％、5％-20％、6％-20％、7％-20％、8％-20％、9％-20％、10％-20％、11％-20％、12％-20％、13％-20％、14％-20％、15％-20％、16％-20％、17％-20％、18％-20％、19％-20％、1％-19％、2％-19％、3％-19％、4％-19％、5％-19％、6％-19％、7％-19％、8％-19％、9％-19％、10％-19％、11％-19％、12％-19％、13％-19％、14％-19％、15％-19％、16％-19％、17％-19％、18％-19％、1％-18％、2％-18％、3％-18％、4％-18％、5％-18％、6％-18％、7％-18％、8％-18％、9％-18％、10％-18％、11％-18％、12％-18％、13％-18％、14％-18％、15％-18％、16％-18％、17％-18％、1％-17％、2％-17％、3％-17％、4％-17％、5％-17％、6％-17％、7％-17％、8％-17％、9％-17％、10％-17％、11％-17％、12％-17％、13％-17％、14％-17％、15％-17％、16％-17％、1％-16％、2％-16％、3％-16％、4％-16％、5％-16％、6％-16％、7％-16％、8％-16％、9％-16%, 10% -16%, 11% -16%, 12% -16%, 13% -16%, 14% -16%, 15% -16%, 1% -15%, 2% -15%, 3% -15%, 4% -15%, 5% -15%, 6% -15%, 7% -15%, 8% -15%, 9% -15%, 10% -15%, 11% -15%, 12% -15%, 13% -15%, 14% -15%, 1% -14%, 2% -14%, 3% -14%, 4% -14%, 5% -14%, 6% -14%, 7% -14%, 8% -14%, 9% -14%, 10% -14%, 11% -14%, 12% -14%, 10% -14%, 13% -14%, 1% -13%, 2% -13%, 3% -13%, 4% -13%, 5% -13%, 6% -13%, 7% -13%, 8% -13%, 9% -13%, 10% -13%, 11% -13%, 12% -13%, 1% -12%, 2% -12%, 3% -12%, 4% -12%, 5% -12%, 6% -12%, 7% -12%, 8% -12%, 9% -12%, 10% -12%, 11% -12%, 1% -11%, 2% -11%, 3% -11%, 4% -11%, 5% -11%, 6% -11%, 7% -11%, 8% -11%, 9% -11%, 10% -11%, 1% -10%, 2% -10%, 3% -10%, 4% -10%, 5% -10%, 6% -10%, 7% -10%, 8% -10%, 9% -10%, 1% -9%, 2% -9%, 3% -9%, 4% -9%, 5% -9%, 6% -9%, 7% -9%, 8% -9%, 1% -8%, 2% -8%, 3% -8%, 4% -8%, 5% -8%, 6% -8%, 7% -8%, 1% -7%, 2% -7%, 3% -7%, 4% -7%, 5% -7%, 6% -7%, 1% -6%, 2% -6%, 3% -6%, 4% -6%, 5% -6%, 1% -5%, 2% -5%, 3% -5%, 4% -5%, 1% -4%, 2% -4%, 3% -4%, 1% -3%, 2% -3% or 1% -2% glycerol.
In some embodiments, the compositions as described herein have a higher ratio than typical yeast products used for fermentation (e.g., fali, ethanol) at temperatures of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid Angel Super/>) Has higher fermentation rate. In a particular embodiment, the compositions as described herein are specific to typical yeast products used for fermentation (e.g., fali, ethanol->Angel Super/>) Has a significantly higher fermentation rate. In another particular embodiment, the compositions as described herein are specific for typical yeast products used for fermentation at a temperature of 36 ℃ and at low pH of the organic acid (e.g., fali, ethanol->Angel Super/>) Has a significantly higher fermentation rate.
In some embodiments, the compositions as described herein have a fermentation rate after fermentation for 24 hours at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid that is greater than typical yeast products used for fermentation (e.g., fali, ethanolAngel Super/>) At least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% higher. In some embodiments, the compositions as described herein have a fermentation rate after fermentation for 24 hours at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid that is greater than typical yeast products used for fermentation (e.g., fa li, ethane ol + >Angel Super/>) Up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% or 50% higher. In some embodiments, the compositions as described herein have a fermentation rate ratio after fermentation for 24 hours at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at a low pH of the organic acid, to typical yeast products used for fermentation (e.g., fa li, ethane ol +>Angel Super) From about 1% -50% (i.e., from about 1% -50%), 2% -50%, 3% -50%, 4% -50%, 5% -50%, 6% -50%, 7% -50%, 8% -50%, 9% -50%, 10% -50%, 11% -50%, 12% -50%, 13% -50%, 14% -50%, 15% -50%, 16% -50%, 17% -50%, 18% -50%, 19% -50%, 20% -50%, 21% -50%, 22% -50%, 23% -50%, 24% -50%, 25% -50%, 26% -50%, 27% -50%, 28% -50%, 2% -49%, 3% -49%, and 4% -49%, 5% -49%, 6% -49%, 7% -49%, 8% -49%, 9% -49%, 10% -49%, 11% -49%, 12% -49%, 13% -49%, 14% -49%, 15% -49%, 16% -49%, 17% -49%, 18% -49%, 19% -49%, 20% -49%, 21% -49%, 22% -49%, 23% -49%, 24% -49%, 25% -49%, 26% -49%, 27% -49%, 28% -49%, 2% -48%, 3% -48%, 4% -48%, 5% -48%, 6% -48%, 7% -48%, 8% -48%, 9% -48%, 10% -48%, 11% -48%, 12% -48%, 13% -48%, 14％-48％、15％-48％、16％-48％、17％-48％、18％-48％、19％-48％、20％-48％、21％-48％、22％-48％、23％-48％、24％-48％、25％-48％、26％-48％、27％-48％、28％-48％、2％-47％、3％-47％、4％-47％、5％-47％、6％-47％、7％-47％、8％-47％、9％-47％、10％-47％、11％-47％、12％-47％、13％-47％、14％-47％、15％-47％、16％-47％、17％-47％、18％-47％、19％-47％、20％-47％、21％-47％、22％-47％、23％-47％、24％-47％、25％-47％、26％-47％、27％-47％、28％-47％、2％-46％、3％-46％、4％-46％、5％-46％、6％-46％、7％-46％、8％-46％、9％-46％、10％-46％、11％-46％、12％-46％、13％-46％、14％-46％、15％-46％、16％-46％、17％-46％、18％-46％、19％-46％、20％-46％、21％-46％、22％-46％、23％-46％、24％-46％、25％-46％、26％-46％、27％-46％、28％-46％、2％-45％、3％-45％、4％-45％、5％-45％、6％-45％、7％-45％、8％-45％、9％-45％、10％-45％、11％-45％、12％-45％、13％-45％、14％-45％、15％-45％、16％-45％、17％-45％、18％-45％、19％-45％、20％-45％、21％-45％、22％-45％、23％-45％、24％-45％、25％-45％、26％-45％、27％-45％、28％-45％、2％-44％、3％-44％、4％-44％、5％-44％、6％-44％、7％-44％、8％-44％、9％-44％、10％-44％、11％-44％、12％-44％、13％-44％、14％-44％、15％-44％、16％-44％、17％-44％、18％-44％、19％-44％、20％-44％、21％-44％、22％-44％、23％-44％、24％-44％、25％-44％、26％-44％、27％-44％、28％-44％、2％-43％、3％-43％、4％-43％、5％-43％、6％-43％、7％-43％、8％-43％、9％-43％、10％-43％、11％-43％、12％-43％、13％-43％、14％-43％、15％-43％、16％-43％、17％-43％、18％-43％、19％-43％、20％-43％、21％-43％、22％-43％、23％-43％、24％-43％、25％-43％、26％-43％、27％-43％、28％-43％、2％-42％、3％-42％、4％-42％、5％-42％、6％-42％、7％-42％、8％-42％、9％-42％、10％-42％、11％-42％、12％-42％、13％-42％、14％-42％、15％-42％、16％-42％、17％-42％、18％-42％、19％-42％、20％-42％、21％-42％、22％-42％、23％-42％、24％-42％、25％-42％、26％-42％、27％-42％、28％-42％、2％-41％、3％-41％、4％-41％、5％-41％、6％-41％、7％-41％、8％-41％、9％-41％、10％-41％、11％-41％、12％-41％、13％-41％、14％-41％、15％-41％、16％-41％、17％-41％、18％-41％、19％-41％、20％-41％、21％-41％、22％-41％、23％-41％、24％-41％、25％-41％、26％-41％、27％-41％、28％-41％、2％-40％、3％-40％、4％-40％、5％-40％、6％-40％、7％-40％、8％-40％、9％-40％、10％-40％、11％-40％、12％-40％、13％-40％、14％-40％、15％-40％、16％-40％、17％-40％、18％-40％、19％-40％、20％-40％、21％-40％、22％-40％、23％-40％、24％-40％、25％-40％、26％-40％、27％-40％、28％-40％、2％-39％、3％-39％、4％-39％、5％-39％、6％-39％、7％-39％、8％-39％、9％-39％、10％-39％、11％-39％、12％-39％、13％-39％、14％-39％、15％-39％、16％-39％、17％-39％、18％-39％、19％-39％、20％-39％、21％-39％、22％-39％、23％-39％、24％-39％、25％-39％、26％-39％、27％-39％、28％-39％、2％-38％、3％-38％、4％-38％、5％-38％、6％-38％、7％-38％、8％-38％、9％-38％、10％-38％、11％-38％、12％-38％、13％-38％、14％-38％、15％-38％、16％-38％、17％-38％、18％-38％、19％-38％、20％-38％、21％-38％、22％-38％、23％-38％、24％-38％、25％-38％、26％-38％、27％-38％、28％-38％、2％-37％、3％-37％、4％-37％、5％-37％、6％-37％、7％-37％、8％-37％、9％-37％、10％-37％、11％-37％、12％-37％、13％-37％、14％-37％、15％-37％、16％-37％、17％-37％、18％-37％、19％-37％、20％-37％、21％-37％、22％-37％、23％-37％、24％-37％、25％-37％、26％-37％、27％-37％、28％-37％、2％-36％、3％-36％、4％-36％、5％-36％、6％-36％、7％-36％、8％-36％、9％-36％、10％-36％、11％-36％、12％-36％、13％-36％、14％-36％、15％-36％、16％-36％、17％-36％、18％-36％、19％-36％、20％-36％、21％-36％、22％-36％、23％-36％、24％-36％、25％-36％、26％-36％、27％-36％、28％-36％、2％-35％、3％-35％、4％-35％、5％-35％、6％-35％、7％-35％、8％-35％、9％-35％、10％-35％、11％-35％、12％-35％、13％-35％、14％-35％、15％-35％、16％-35％、17％-35％、18％-35％、19％-35％、20％-35％、21％-35％、22％-35％、23％-35％、24％-35％、25％-35％、26％-35％、27％-35％、28％-35％、2％-34％、3％-34％、4％-34％、5％-34％、6％-34％、7％-34％、8％-34％、9％-34％、10％-34％、11％-34％、12％-34％、13％-34％、14％-34％、15％-34％、16％-34％、17％-34％、18％-34％、19％-34％、20％-34％、21％-34％、22％-34％、23％-34％、24％-34％、25％-34％、26％-34％、27％-34％、28％-34％、2％-33％、3％-33％、4％-33％、5％-33％、6％-33％、7％-33％、8％-33％、9％-33％、10％-33％、11％-33％、12％-33％、13％-33％、14％-33％、15％-33％、16％-33％、17％-33％、18％-33％、19％-33％、20％-33％、21％-33％、22％-33％、23％-33％、24％-33％、25％-33％、26％-33％、27％-33％、28％-33％、2％-32％、3％-32％、4％-32％、5％-32％、6％-32％、7％-32％、8％-32％、9％-32％、10％-32％、11％-32％、12％-32％、13％-32％、14％-32％、15％-32％、16％-32％、17％-32％、18％-32％、19％-32％、20％-32％、21％-32％、22％-32％、23％-32％、24％-32％、25％-32％、26％-32％、27％-32％、28％-32％、2％-31％、3％-31％、4％-31％、5％-31％、6％-31％、7％-31％、8％-31％、9％-31％、10％-31％、11％-31％、12％-31％、13％-31％、14％-31％、15％-31％、16％-31％、17％-31％、18％-31％、19％-31％、20％-31％、21％-31％、22％-31％、23％-31％、24％-31％、25％-31％、26％-31％、27％-31％、28％-31％、2％-30％、3％-30％、4％-30％、5％-30％、6％-30％、7％-30％、8％-30％、9％-30％、10％-30％、11％-30％、12％-30％、13％-30％、14％-30％、15％-30％、16％-30％、17％-30％、18％-30％、19％-30％、20％-30％、21％-30％、22％-30％、23％-30％、24％-30％、25％-30％、26％-30％、27％-30％、28％-30％、29％-30％、1％-29％、2％-29％、3％-29％、4％-29％、％-29％、6％-29％、7％-29％、8％-29％、9％-29％、10％-29％、11％-29％、12％-29％、13％-29％、14％-29％、15％-29％、16％-29％、17％-29％、18％-29％、19％-29％、20％-29％、21％-29％、22％-29％、23％-29％、24％-29％、25％-29％、26％-29％、27％-29％、28％-29％、1％-28％、2％-28％、3％-28％、4％-28％、5％-28％、6％-28％、7％-28％、8％-28％、9％-28％、10％-28％、11％-28％、12％-28％、13％-28％、14％-28％、15％-28％、16％-28％、17％-28％、18％-28％、19％-28％、20％-28％、21％-28％、22％-28％、23％-28％、24％-28％、25％-28％、26％-28％、27％-28％、1％-27％、2％-27％、3％-27％、4％-27％、5％-27％、6％-27％、7％-27％、8％-27％、9％-27％、10％-27％、11％-27％、12％-27％、13％-27％、14％-27％、15％-27％、16％-27％、17％-27％、18％-27％、19％-27％、20％-27％、21％-27％、22％-27％、23％-27％、24％-27％、25％-27％、26％-27％、1％-26％、2％-26％、3％-26％、4％-26％、5％-26％、6％-26％、7％-26％、8％-26％、9％-26％、10％-26％、11％-26％、12％-26％、13％-26％、14％-26％、15％-26％、16％-26％、17％-26％、18％-26％、19％-26％、20％-26％、21％-26％、22％-26％、23％-26％、24％-26％、25％-26％、1％-25％、2％-25％、3％-25％、4％-25％、5％-25％、6％-25％、7％-25％、8％-25％、9％-25％、10％-25％、11％-25％、12％-25％、13％-25％、14％-25％、15％-25％、16％-25％、％-25％、18％-25％、19％-25％、20％-25％、21％-25％、22％-25％、23％-25％、24％-25％、1％-24％、2％-24％、3％-24％、4％-24％、5％-24％、6％-24％、7％-24％、8％-24％、9％-24％、10％-24％、11％-24％、12％-24％、13％-24％、14％-24％、15％-24％、16％-24％、17％-24％、18％-24％、19％-24％、20％-24％、21％-24％、22％-24％、23％-24％、1％-23％、2％-23％、3％-23％、4％-23％、5％-23％、6％-23％、7％-23％、8％-23％、9％-23％、10％-23％、11％-23％、12％-23％、13％-23％、14％-23％、15％-23％、16％-23％、17％-23％、18％-23％、19％-23％、20％-23％、21％-23％、22％-23％、1％-22％、2％-22％、3％-22％、4％-22％、5％-22％、6％-22％、7％-22％、8％-22％、9％-22％、10％-22％、11％-22％、12％-22％、13％-22％、14％-22％、15％-22％、16％-22％、17％-22％、18％-22％、19％-22％、20％-22％、21％-22％、1％-21％、2％-21％、3％-21％、4％-21％、5％-21％、6％-21％、7％-21％、8％-21％、9％-21％、10％-21％、11％-21％、12％-21％、13％-21％、14％-21％、15％-21％、16％-21％、17％-21％、18％-21％、19％-21％、20％-21％、1％-20％、2％-20％、3％-20％、4％-20％、5％-20％、6％-20％、7％-20％、8％-20％、9％-20％、10％-20％、11％-20％、12％-20％、13％-20％、14％-20％、15％-20％、16％-20％、17％-20％、18％-20％、19％-20％、1％-19％、2％-19％、3％-19％、4％-19％、5％-19％、6％-19％、7％-19％、8％-19％、9％-19％、10％-19％、11％-19％、12％-19％、13％-19％、14％-19％、15％-19％、16％-19％、17％-19％、18％-19％、1％-18％、2％-18％、3％-18％,4％-18％、5％-18％、6％-18％、7％-18％、8％-18％、9％-18％、10％-18％、11％-18％、12％-18％、13％-18％、14％-18％、15％-18％、16％-18％、17％-18％、1％-17％、2％-17％、3％-17％、4％-17％、5％-17％、6％-17％、7％-17％、8％-17％、9％-17％、10％-17％、11％-17％、12％-17％、13％-17％、14％-17％、15％-17％、16％-17％、1％-16％、2％-16％、3％-16％、4％-16％、5％-16％、6％-16％、7％-16％、8％-16％、9％-16％、10％-16％、11％-16％、12％-16％、13％-16％、14％-16％、15％-16％、1％-15％、2％-15％、3％-15％、4％-15％、5％-15％、6％-15％、7％-15％、8％-15％、9％-15％、10％-15％、11％-15％、12％-15％、13％-15％、14％-15％、1％-14％、2％-14％、3％-14％、4％-14％、5％-14％、6％-14％、7％-14％、8％-14％、9％-14％、10％-14％、11％-14％、12％-14％、13％-14％、1％-13％、2％-13％、3％-13％、4％-13％、5％-13％、6％-13％、7％-13％、8％-13％、9％-13％、10％-13％、11％-13％、12％-13％、1％-12％、2％-12％、3％-12％、4％-12％、5％-12％、6％-12％、7％-12％、8％-12％、9％-12％、10％-12％、11％-12％、1％-11％、2％-11％、3％-11％、4％-11％、5％-11％、6％-11％、7％-11％、8％-11％、9％-11％、10％-11％、1％-10％、2％-10％、3％-10％、4％-10％、5％-10％、6％-10％、7％-10％、8％-10％、9％-10％、1％-9％、2％-9％、3％-9％、4％-9％、5％-9％、6％-9％、7％-9％、8％-9％、1％-8％、2％-8％、3％-8％、4％-8％、5％-8％、6％-8％、7％-8％、1％-7％、2％-7％、3％-7％、4％-7％、5％-7％、6％-7％、1％-6％、2％-6％、3％-6％、4％-6％、5％-6％、1％-5％、2％-5％、3％-5％、4％-5％、1％-4％、2％-4％、3％-4％、1％-3％、2％-3% or 1% -2%.
In some embodiments, the compositions as described herein have one or more of the defined features described herein. In some embodiments, the compositions as described herein have one or more of the defined features described herein at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at low pH of the organic acid. In some embodiments, the compositions as described herein have one or more of the defined characteristics described herein at a temperature of 20 ℃ to 40 ℃ (preferably 32 ℃ to 36 ℃) and at normal pH without an organic acid. For example, in some embodiments, a composition as described herein is more effective than a typical yeast product used for fermentation (e.g., fali, ethanolAngel Super/>) Has higher ethanol yield, lower glycerol yield, higher fermentation rate, higher temperature tolerance and higher organic acid tolerance; in some embodiments, the composition as described herein is combined with a typical yeast product for fermentation (e.g., fali, ethanol->Angel Super/>) Lower levels of glycerol are produced and significantly higher ethanol yields are achieved than with higher fermentation rates; in some embodiments, the composition as described herein is combined with a typical yeast product for fermentation (e.g., fali, ethanol- >Angel Super/>) Has a higher fermentation rate and produces lower levels of glycerol than would otherwise be produced.
4. Ethanol production
Described herein are methods for producing ethanol from a substrate by contacting the substrate with a fermenting organism or a composition comprising a fermenting organism. The fermenting organism is selected from the group consisting of yeast strains and derivatives thereof described herein. Saccharomyces cerevisiae (Saccharomyces cerevisiae) Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or fermenting organisms having substantially the same properties as the yeast strains described herein or derivatives of the yeast strains described herein with defined characteristics may be used in the methods described herein. Also described herein are fermentation products comprising any of the yeasts described herein. Further described herein are fermentation products obtained by the methods described herein. The fermentation products may include, but are not limited to, fuel ethanol, industrial ethanol, potable ethanol, bioethanol, fermented foods (such as alcoholic beverages, cultured milk and yogurt, wine, beer, cider, indonesia fermented soybean, miso, kimchi, pickled vegetables, and fermented sausage).
The fermentation is carried out in a fermentation medium. The fermentation medium comprises a fermentation substrate, i.e. a carbohydrate source such as biomass, which is metabolized by the fermenting organism. The fermentation medium may comprise nutrients of the fermenting organism. Nutrients are widely used in fermentation processes and include nitrogen sources, vitamins, minerals, or combinations thereof.
In one embodiment, the strain or derivative or composition as described herein is incubated with a substrate comprising fermentable sugars from biomass (e.g., plant biomass from forests and/or from agricultural or food processing products and/or byproducts) that constitutes a substantial carbon source for the production of the molecule of interest. The strain or derivative is incubated with the substrate under conditions that allow for fermentation of the fermentable sugar. The fermentable sugar may be glucose, galactose, maltose, fructose, sucrose, mannose, or a combination thereof. Typically, the fermentable sugars are glucose and sucrose. The source of fermentable sugars in the substrate may be any source containing fermentable sugars. For example, the fermentable sugars in the substrate may be from any one or more of the following sources: hydrolyzed starch, hydrolyzed cellulose, molasses (from sugar cane or sugar beet), sugar cane juice, agave, sugar beet juice, grape juice, fruit juice, glucose, hydrolyzed maltodextrin, raw sugar juice, galactose, sucrose, any other form of fermentable sugar or combinations thereof. Starch may be obtained from any starch-rich crop. Examples of starch-rich crops include, but are not limited to, corn, wheat, barley, tapioca, sorghum, sweet potato, millet, rice, or any other starch-rich crop. In preparing the substrate, the crop is typically crushed and mixed with water and a hydrolase under conditions that cause the starch to hydrolyze and release fermentable sugars (e.g., glucose). Typical enzymes for hydrolyzing starch include alpha-amylase, amyloglucosidase, pullulanase, beta-amylase, glucoamylase, or mixtures thereof.
Typically, fermenting organisms such as yeast (including Saccharomyces cerevisiae) require a sufficient nitrogen source to reproduce and ferment. Many nitrogen sources may be used and such nitrogen sources are well known in the art. The nitrogen source may be organic, such as urea or corn steep liquor, or inorganic, such as ammonia or ammonium hydroxide or ammonium salts.
In a particular embodiment, the biomass may comprise or be derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or combinations thereof. In a particular embodiment, the substrate is provided in the form of corn slurry or Synthetic Corn Medium (SCM). Methods of preparing mashed corn are known in the art and are described, for example, in Thomas et al, (2001) Journal of Applied Microbiology,90,819-828. Methods of preparing substrates that function similarly to SCM are known in the art and are described, for example, in U.S. patent No. 10,106,823, which is incorporated herein by reference in its entirety. Methods of preparing starch-based substrates are also described, for example, in PCT publication No. 2006/113683 or U.S. patent publication No. 2007/0014905.
The sugar content of the fermentation medium can be adjusted to be as high as possible while ensuring as rapid and complete conversion of the sugar into ethanol as possible. Preferably, the yeast converts all of the sugars in the medium to ethanol and the total yield of consumed sugars to ethanol is as high as possible and, therefore, minimal by-products (such as glycerol) are produced during fermentation.
The fermentation is carried out at a temperature that allows fermenting the fermentable sugar. Generally, the higher the fermentation temperature, the more economical the industrial process. Typically, the fermentation is carried out at a temperature of 25-42 ℃ (i.e. from 25 ℃ to 42 ℃). Suitable temperature ranges include 25-41 ℃, 26-40 ℃, 27-40 ℃, 28-40 ℃, 29-40 ℃, 30-40 ℃, 25-39 ℃, 26-39 ℃, 27-39 ℃, 28-39 ℃, 29-39 ℃, 30-39 ℃, 31-39 ℃, 32-39 ℃, 33-39 ℃, 25-38 ℃, 26-38 ℃, 27-38 ℃, 28-38 ℃, 29-38 ℃, 30-38 ℃, 31-38 ℃, 32-38 ℃, 33-38 ℃, 25-37 ℃, 26-37 ℃, 27-37 ℃, 28-37 ℃, 29-37 ℃, 30-37 ℃, 31-37 ℃, 32-37 ℃, 33-37 ℃, 25-36 ℃, 26-36 ℃, 27-36 ℃, 28-36 ℃, 29-36 ℃, 30-36 ℃, 31-36 ℃, 32-36 ℃, 33-36 ℃, 25-35 ℃, 26-35 ℃, 27-35 ℃, 28-35 ℃, 29-35 ℃, 30-35 ℃, 31-35 ℃, 32-35 ℃, or 33-35 ℃.
Methods of fermentation and distillation are known in the art and are described, for example, in WO 2006/113683 or US 2007/0014905. In particular, ethanol production as described herein may be performed using simultaneous saccharification and fermentation ("SSF"), batch fermentation, or continuous fermentation.
a. Synchronous saccharification and fermentation ('SSF')
SSF is widely used in industrial scale fermentation product production processes, particularly ethanol production processes. When SSF is performed, the saccharification step and fermentation step are performed simultaneously. Saccharification does not have a hold stage, which means that the fermenting organism (e.g. yeast) and enzyme can be added together. However, separate addition of fermenting organisms and enzymes (i.e. Step Hydrolytic Fermentation (SHF)) is also contemplated. SSF may be performed at a temperature of 25 ℃ to 40 ℃, such as 28 ℃ to 35 ℃, such as 30 ℃ to 34 ℃, preferably about 32 ℃. In one embodiment, the fermentation lasts from 6 to 120 hours, in particular from 24 to 96 hours, and preferably about 50 hours. In one embodiment, the pH is from about 3.0 to about 6.0, preferably from about 4.0 to about 5.0.
After fermentation (e.g., SSF), ethanol may be separated from spent fermentation media or beer. Beer can be rectified or distilled to recover/extract the desired fermentation products (i.e., ethanol and higher alcohols). Alternatively, the desired fermentation product (i.e., ethanol) may be extracted from the fermentation medium by microfiltration or membrane filtration techniques known in the art. The fermentation product (i.e., ethanol) may also be recovered by steam stripping or other methods known in the art.
In some embodiments, the ethanol is not recovered/extracted from the fermentation medium or beer, such as for use in producing an alcoholic beverage.
b. Batch fermentation
Batch fermentation is fermentation performed in separate batches. Batch fermentation is a process in which a fermentation medium is provided starting from a fermenter which is inoculated with the desired microorganism (i.e. yeast, yeast product) and the fermentation process is continued until a predetermined condition is reached, typically the consumption of substrate in the fermentation medium and the resulting cessation of ethanol production by the consumption. Once the process is complete, the product is removed from the fermenter and the fermenter is sterilized prior to the next fermentation. The contents are then the final product (e.g. wine) or may be rectified/distilled (e.g. fuel ethanol and whiskey).
Fed-batch processes may also be used. A fed-batch process is a fermentation in which a portion of the fermentation medium is provided from the beginning of the fermentation process with the addition of an inoculum and at some point in time after the start of the fermentation, the feed enters the fermenter at a rate predetermined or determined by the conditions in the fermenter; until the maximum volume is reached. The feed may or may not have the same composition as the initial fermentation medium. The contents are then the final product (e.g. wine) or may be rectified/distilled (e.g. fuel ethanol and whiskey).
c. Continuous fermentation
Continuous fermentation allows for long fermentation without the need for batch fermentation. A continuous fermentation process is a process in which fresh growth medium is continuously fed into a fermenter and fermentation broth is withdrawn from the fermenter at the same rate while the volume in the fermenter is kept constant. The contents are then the final product (e.g. wine) or may be rectified/distilled (e.g. fuel ethanol and whiskey).
5. Ethanol
As described herein, the ethanol produced by the yeast and its derivatives may be fuel ethanol, industrial ethanol, and/or potable ethanol. Fuel ethanol, industrial ethanol, and potable ethanol can be produced from biomass containing starch, including starch in cereals (e.g., corn, wheat, rice, sorghum/milo, barley, etc.), and starch in tubers and root vegetables (e.g., potato, tapioca, etc.); and the vegetative parts of plants containing sucrose, glucose and fructose (e.g., sugar cane, sweet sorghum, sugar beet, agave, etc.); and fruits and berries from plants containing sucrose, glucose and fructose (e.g., grape, orange, peach, cherry, etc.).
Fuel ethanol and industrial ethanol can also be produced from plant biomass containing cellulose and hemicellulose, such as cereal crop residues (e.g., wheat and rice straw, corn stover, corn cobs, etc.), from corn fiber, from so-called energy crops (e.g., switchgrass and poplar), from wood material waste (including saw mill residues (e.g., sawdust and wood chips)), from pulp and paper manufacturing residues, and waste paper and board.
a. Ethanol fuel
The manufacture of fuel ethanol is for an internal combustion engine and may be made as anhydrous or aqueous fuel ethanol. The anhydrous fuel ethanol may be mixed with gasoline to form an ethanol/gasoline mixture or with diesel to form an ethanol/diesel mixture. The aqueous fuel ethanol can be used directly as a fuel for an internal combustion engine.
b. Industrial ethanol
The manufacture of industrial ethanol is used in a variety of applications, including as a solvent for pharmaceuticals, cosmetics, detergents, household cleaners and disinfectants, paints and inks; and as chemical intermediates for the manufacture of ethyl acetate, ethyl acrylate, polyethylene, acetic acid and other organic molecules of industrial importance.
c. Drinking ethanol
The production of potable ethanol is for human consumption, including ethanol in wine, beer, cider, sake, honey wine, kang Pucha, and distilled spirits, including whiskey, bouillon, coffee, chinese spirits, white spirits, and the like.
6. Yeast strain production
a. Directional mating
Provided herein are methods of producing derivatives of yeast (Saccharomyces) strains as described herein. The method may include providing a first yeast strain selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927 and Y1929 and a second yeast strain of any yeast strain, such as a yeast strain in the Saccharomyces narrow (Saccharomyces sensu stricto) clade, such as a Saccharomyces cerevisiae (Saccharomyces cerevisiae) strain. The second strain may also be any yeast strain described herein. The method may further comprise inducing sporulation of the first yeast strain and the second yeast strain. The method may further comprise screening and selecting spores from the first yeast strain and the second yeast strain. Additionally, the method may include hybridizing selected spores of the first yeast strain with selected spores of the second yeast strain, and screening or selecting the derivative strain. The method may comprise screening or selecting spores that exhibit one or more defined characteristics of a yeast (Saccharomyces) strain as described herein. The method further may comprise screening or selecting hybrids that exhibit one or more defined characteristics of a yeast (Saccharomyces) strain as described herein. Fig. 1A provides an example of directional mating. Thus, the parents (i.e., the "a" and "a" haploid donors) that produce the hybrid are known. In one embodiment, the yeast strains and derivatives thereof as described herein are made by the method shown in fig. 1. In one embodiment, the yeast (Saccharomyces) strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 are derived from one or more different yeast (Saccharomyces) strains by the process shown in FIG. 1 (e.g., by the process shown in FIG. 1A), and thus one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 are products of the directed mating. Methods of directional mating are known in the art and are described in U.S. patent No. 10,308,963 and U.S. patent No. 10,106,823, which are incorporated herein by reference.
b. Large scale mating
Provided herein are methods of producing derivatives of yeast (Saccharomyces) strains as described herein. The method may include providing a first yeast strain selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929, and one or more additional yeast strains, such as a Saccharomyces cerevisiae strain in the narrow (Saccharomyces sensu stricto) clade, such as a Saccharomyces cerevisiae (Saccharomyces cerevisiae) strain, as any of the yeast strains. The one or more additional yeast strains may also be any yeast strain as described herein. The method may further comprise inducing sporulation of the first yeast strain and the one or more additional yeast strains. The method may further comprise mixing all spores to allow hybridization of the spores and screening or selecting the derivative strain. The method may comprise screening or selecting hybrids that exhibit one or more defined characteristics of a yeast strain as described herein. Fig. 1B provides an example of a large-scale mating. Thus, the parents that produce the hybrid (i.e., the "a" and "a" haploid donors) are unknown. In one embodiment, the yeast strains and derivatives thereof as described herein are derived from the process shown in fig. 1. In one embodiment, the yeast strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 are derived from one or more different yeast(s) (Saccharomyces) strains by the process shown in fig. 1 (e.g., by the process shown in fig. 1B), and thus one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 are products of large scale mating.
7. Examples
The foregoing may be better understood by reference to the following examples which are intended for the purpose of illustration only and are not intended to limit the scope of the present invention. The present disclosure has various aspects and embodiments, illustrated by the attached non-limiting examples.
Example 1
Materials and methods
Synthetic Corn Media (SCM) was used in the following examples. SCM mimics corn steep liquor, which is commonly used in industrial bioethanol production. SCM is intended to reflect the chemical composition of yellow dent corn #2 in terms of sugar and nutrient availability. SCM reflects a high gravity model corn system with a solids content of 32% and a starch content of about 72% and a final ethanol yield of 13-14% (w/v). The synthetic nature of the medium allows for stringent replication of the test conditions for each experiment.
The medium is used to monitor the performance of yeast in a Simultaneous Saccharification and Fermentation (SSF) process, wherein starch in the medium is broken down into glucose by the use of glucoamylase (saccharification), and glucose is used simultaneously (simultaneously) by the yeast for the production of ethanol (fermentation).
Preparation of SCM. 1L of SCM complete medium was prepared by adding 500g of water to a 2L beaker. The ingredients in table 1 were added to the beaker and less than 100 milliliters of water was used in total. The solution was stirred at 300rpm and gradually heated to 75 ℃. Once the solution reached 75 ℃, it was removed from the heat and cooled to ambient temperature while mixing. During mixing, 100.0ml of the 10×scm vitamin mixture (table 2) and 10.0ml of the 100×scm mineral mixture (table 3) were added to the solution. Then, while stirring, 25.0mL of SCM amino acid slurry (Table 4), 1.2G of urea (550 ppm nitrogen equivalent), 0.400mL of a Virginia mycin stock solution (1.0 ppm) and 0.1G of penicillin G powder (100 ppm) were added to the solution. The solution was transferred to a 1 liter volumetric flask and made up with water to 1 liter. The pH of the solution was adjusted to 5.0-5.2 with 28% sulfuric acid or 28% ammonium/potassium hydroxide. The solution was mixed by inversion and further stirring at 300-450 rpm. Representative samples of the solutions were collected for specific gravity measurement and HPLC analysis.
Table 1:1 liter of SCM complete medium composition
| Composition of the components | Specification of specification | Gram/liter SCM |
| Maltodextrin M100 | 100.0% purity, 6% moisture | 295.10 |
| Dipotassium hydrogen phosphate | 98.0% purity, 1.0% moisture | 1.16 |
| Monopotassium phosphate | 99.0% purity, 0.2% moisture | 10.22 |
| Magnesium sulfate heptahydrate | 99.0% purity, 51.1% moisture | 5.54 |
| Sodium chloride | 99.0% purity | 0.30 |
| Calcium chloride dihydrate | 99.0% purity, 24.5% moisture | 0.43 |
Table 2:1 liter of a 10 XSCM vitamin mixture
Table 3:100 ml 100 XSCM mineral mixture composition
| Composition of the components | Specification of specification | Mg/100 ml SCM vitamin mixture |
| Ferric chloride hexahydrate (III) | 97.0% purity, 40% moisture | 578.7 |
| Zinc sulfate heptahydrate | 99.0% purity, 43.8% moisture | 240.5 |
| Manganese sulfate monohydrate (II) | 99.0% purity, 10.7% moisture | 60.1 |
| Pentahydrate copper sulfate (II) | 99.0% purity, 36% moisture | 62.0 |
Table 4: composition of 200 ml SCM amino acid slurry
For each yeast strain, 100.00±0.10mL of SCM was added to each test sample container. Each test sample was tested in at least three replicates. All test sample containers with SCM were incubated at 150rpm for 1 to 2 hours at 32 ℃ to equilibrate to temperature. The specific gravity of each sample container was measured with a densitometer. Samples were removed from the sample containers for HPLC analysis. The sample container was rotated to ensure all cells were suspended and 1.9mL of sample container medium was placed into a 2mL microcentrifuge tube. The sample was centrifuged at 3000rpm for 10 minutes. The supernatant was poured into a syringe with a pre-installed 0.2 μm filter and the filtrate was collected into a glass HPLC vial. HPLC vials with samples can be stored at +4 ℃ for up to 1 month or analyzed immediately prior to HPLC analysis.
Isolating the Saccharomyces cerevisiae strain. Methods of isolating yeast strains from agar plates are known in the art.
And (5) culturing yeast. 2.5mL of yeast culture (i.e., yeast in common yeast growth medium; e.g., glucose yeast peptone medium (GYP)) was inoculated into a sample vessel with SCM to 1.0X10 6 Final cell concentration of individual cells/mL.
Yeast slants or colonies on plates. The whole 10. Mu.L of cells on the loop were inoculated in 50mL of 2% sterile GYP. The cultures were incubated overnight at 32℃or 36℃and 150rpm to give 1X 10 8 Cell concentration of individual cells/mL. 2.5mL of culture was inoculated into a sample vessel with SCM to 2.5X10 6 Final cell concentration of individual cells/mL. Immediately after all yeast samples were added, 10-fold dilution of glucoamylase was added to the sample container. In the case of 140. Mu. LSpirizymeFuel HS addition, 224. Mu.L was added to the Spirizyme Ultra. If an alternative glucoamylase is used, the enzyme dosage must be properly adjusted to 100 ml on a 32% dry corn solids basis. Typically, the dose is expressed in% w/w units, so that the SCM solution density needs to be considered.
The sample container is closed with a airlock cover. The pressurized air lock was filled with 9 ml of sterile water. The best approach is to place the tip of the pipette into the air lock and dispense as close as possible to the air bubble. The sample containers were incubated at 32℃or 36℃at 150rpm and 80% humidity for 50 hours. The weight of the sample containers was measured before incubation and at the following time points after the start of incubation: 18. 24, 44 and 50 hours. After 50 hours of incubation, the sample containers were removed from the incubator and sampled using HPLC analysis as described above. The specific gravity of the sample container was measured with a densitometer. The pH of the remaining sample in the sample container is then measured. After pH measurement, yeast cell concentration and viability were measured on a hemocytometer using methylene blue.
And (5) carrying out statistical analysis. Analysis of variance (ANOVA) was performed on the final mass loss, ethanol, glycerol and glucose values, and 95% confidence intervals (95% ci) were calculated for each analyte. Preferably, a SAS JMP procedure is used, for example.
Example 2
Description of strains
Representative samples of the yeast (Saccharomyces) strains as described herein, and the strains that have been deposited with NRRL accession number Y-68003 (Y1912), NRRL accession number Y-68004 (Y1913), NRRL accession number Y-68005 (Y1914), NRRL accession number Y-68006 (Y1919), NRRL accession number Y-68007 (Y1923), NRRL accession number Y-68008 (Y1927), and NRRL accession number Y-68009 (Y1929), were observed to have the following characteristics at normal pH without organic acid or at low pH with organic acid based on 4 replicates at 32℃or 36 ℃. Y1609 is shown for comparison and analysis is performed under similar conditions.
Significant and advantageous features of the yeast strains as described herein include increased metabolic ethanol increase, decreased metabolic glycerol and increased initial fermentation rate compared to controls. Advantageous fermentation characteristics of the yeast strains as described herein are provided in the examples below.
Example 3
Ethanol production, glycerol production and fermentation rates
Ethanol production, glycerol production and fermentation rates of yeast strains as described herein were evaluated in synthetic corn starch medium (SCM) at 32 ℃ and pH 5.2 (table 5 and fig. 2). A control (Y1609) is shown for comparison.
Table 5: defined characteristics relative to control at fermentation temperature of 32℃and pH 5.2
Ethanol production, glycerol production and fermentation rates of parent yeast strains as described herein were assessed in SCM exogenously added with 1% w/v lactic acid and 0.05% w/v acetic acid at 32 ℃ and pH 4.0 (table 6 and figure 3). A control (Y1609) is shown for comparison.
Table 6: defined characteristics relative to control at fermentation temperature of 32 ℃ and pH 4.0 with organic acid added
Ethanol production, glycerol production and fermentation rates of parent yeast strains as described herein were assessed in SCM without organic acid at 36 ℃ and pH 5.2 (table 7 and fig. 4). A control (Y1609) is shown for comparison.
Table 7: defined characteristics relative to control at fermentation temperature of 36 ℃ and pH 5.2
Ethanol production, glycerol production and fermentation rates of parent yeast strains as described herein were assessed in SCM exogenously added with 1% w/v lactic acid and 0.05% w/v acetic acid at 36 ℃ and pH 4.0 (table 8 and fig. 5). A control (Y1609) is shown for comparison.
Table 8: defined characteristics relative to control at fermentation temperature of 36 ℃ and pH 4.0 with organic acid added
The foregoing description of the specific aspects will so fully reveal the general nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific aspects without undue experimentation, without departing from the general concept of the present disclosure. Accordingly, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.
The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary aspects, but should be defined only in accordance with the following claims and their equivalents.
All publications, patents, patent applications, and/or other documents cited in this application are incorporated by reference in their entirety for all purposes as if each individual publication, patent application, and/or other document were individually indicated to be incorporated by reference for all purposes.
For completeness, aspects of the invention are listed in the following numbered clauses:
clause 1. A yeast (Saccharomyces) strain selected from the group consisting of: (a) Yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003; (b) Yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004; (c) Yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005; (d) Yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006; (e) Yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit No. Y-68007; (f) Yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009.
Clause 2. A derivative of a yeast (Saccharomyces) strain selected from the group consisting of: (a) Yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003; (b) Yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004; (c) Yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005; (d) Yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006; (e) Yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit No. Y-68007; (f) Yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009.
Clause 3. The yeast strain of clause 1 or the derivative of clause 2, wherein the yeast strain or the derivative comprises one or more defined features selected from the group consisting of: (a) Under the same fermentation conditions, higher ethanol yields than yeast (Saccharomyces) strain Y1609; (b) Has increased temperature tolerance compared to yeast (Saccharomyces) strain Y1609 at a temperature of 32℃to 36 ℃; (c) Lower glycerol yields than yeast (Saccharomyces) strain Y1609 under the same fermentation conditions; (d) Has increased organic acid tolerance compared to yeast (Saccharomyces) strain Y1609 in the presence of an organic acid at a pH of from about 4.0 to about 5.2; and (e) has an increased fermentation rate compared to yeast (Saccharomyces) strain Y1609 under the same fermentation conditions.
Clause 4. The yeast strain or the derivative of any of the preceding clauses, wherein the yeast strain or the derivative produces at least about 0.5% more ethanol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
Clause 5 the yeast strain or the derivative of any of the preceding clauses, wherein the yeast strain or the derivative produces at least about 2% less glycerol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
Clause 6 the yeast strain or the derivative of any of the preceding clauses, wherein the yeast strain or the derivative has a fermentation rate at least about 2% higher than saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609 after 24 hours of fermentation.
Clause 7. The yeast strain of any of clauses 3-6, or the derivative, wherein the temperature is 32 ℃.
Clause 8. The yeast strain of clause 3 or the derivative, wherein the yeast strain or the derivative produces at least about 5% more ethanol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
Clause 9. The yeast strain of clause 3 or the derivative, wherein the yeast strain or the derivative produces at least about 3% less glycerol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
Clause 10. The yeast strain of clause 3 or the derivative, wherein the yeast strain or the derivative has a fermentation rate after 24 hours of fermentation that is at least about 10% higher than that of saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
Clause 11. The yeast strain or the derivative of any of clauses 3 or 8-11, wherein the temperature is 36 ℃.
The yeast strain of any one of clauses 3-11, or the derivative, wherein the organic acid comprises lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, or a combination thereof.
Clause 13. A method of producing a derivative of the yeast (Saccharomyces) strain of clause 2, comprising: (a) providing: (i) A first yeast strain, wherein the first yeast strain is selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; and (ii) a second yeast strain, wherein the second yeast strain is in a yeast narrow strain (Saccharomyces sensu stricto) clade; (b) Inducing sporulation of the first yeast strain and the second yeast strain; (c) Screening and selecting spores from the first yeast strain and spores from the second yeast strain; (d) Hybridizing selected spores of the first yeast strain with selected spores of the second yeast strain; and (e) screening or selecting the derivative strain.
The method of clause 14, clause 13, wherein step (c) comprises screening or selecting spores that exhibit one or more defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.
The method of clause 15, clause 13, wherein step (e) comprises screening or selecting hybrids that exhibit one or more defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.
Clause 16. A method of producing a derivative of the yeast (Saccharomyces) strain of clause 2, comprising: (a) providing: (i) A first yeast strain, wherein the first yeast strain is selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; and (ii) one or more additional yeast strains in the evolution of yeast narrow-sense clades (Saccharomyces sensu stricto); (b) Inducing sporulation of the first yeast strain and the one or more additional yeast strains to produce spores; (c) Mixing all spores of step (b) to allow hybridization of the spores; and (d) screening or selecting the derivative strain.
The method of clause 17, clause 16, wherein step (d) comprises screening or selecting hybrids that exhibit one or more of the defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929.
Clause 18. Mutant yeasts of the yeast strain of clause 1 or the derivative of clause 2.
Clause 19. A method of producing the mutant yeast of clause 18, wherein the mutant yeast is mutated by contacting the yeast strain with a mutagen.
The method of clause 20, clause 19, wherein the mutagen is Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine 2 (ICR-170), or nitrogen mustard.
Clause 21. A method of producing the mutant yeast of clause 18, wherein the mutant yeast is mutated by contacting the derivative with a mutagen.
The method of clause 22, clause 21, wherein the mutagen is Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine 2 (ICR-170), or nitrogen mustard.
Clause 23. Evolved yeast of the yeast strain of clause 1 or the derivative of clause 2.
Clause 24. A method of producing the evolved yeast of clause 23, wherein the evolution is induced by applying a selection pressure to the yeast strain.
Clause 25. A method of producing the evolved yeast of clause 23, wherein the evolution is induced by applying a selection pressure to the derivative.
Clause 26. Genetically modified yeasts of the yeast strain of clause 1 or the derivative of clause 2.
The genetically modified yeast of clause 27, 26, wherein the nucleic acid sequence of the genetically modified yeast is altered using genetic editing.
Clause 28. Recombinant yeast of the yeast strain of clause 1 or the derivative of clause 2.
The recombinant yeast of clause 29, clause 28, wherein the recombinant yeast comprises a modification to inhibit gene expression, enhance gene expression, introduce a gene, or delete a gene.
Clause 30. A method of producing ethanol from a substrate by contacting the substrate with a fermenting organism, wherein the fermenting organism is selected from the group consisting of: (a) A yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003, or a derivative thereof; (b) A yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004, or a derivative thereof; (c) A yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005, or a derivative thereof; (d) A yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006, or a derivative thereof; (e) A yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit number Y-68007, or a derivative thereof; (f) A yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008, or a derivative thereof; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009, or a derivative thereof.
The method of clause 31, clause 30, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or a combination thereof.
The method of clause 32, clause 30, wherein the yeast strain comprises one or more defined features selected from the group consisting of: (a) Under the same fermentation conditions, higher ethanol yields than yeast (Saccharomyces) strain Y1609; (b) Has increased temperature tolerance compared to yeast (Saccharomyces) strain Y1609 at a temperature of 32℃to 36 ℃; (c) Lower glycerol yields than yeast (Saccharomyces) strain Y1609 under the same fermentation conditions; (d) Has increased organic acid tolerance compared to yeast (Saccharomyces) strain Y1609 in the presence of an organic acid at a pH of from about 4.0 to about 5.2; and (e) has an increased fermentation rate compared to yeast (Saccharomyces) strain Y1609 under the same fermentation conditions.
Clause 33, the method of clause 30, wherein the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.
Clause 34. The method of clause 30, wherein the ethanol is produced using starch.
The method of clause 35, clause 34, wherein Simultaneous Saccharification and Fermentation (SSF) or continuous fermentation is used to produce the ethanol.
The method of clause 36, clause 30, wherein the ethanol is produced using a sugar.
The method of clause 37, clause 36, wherein batch fermentation or continuous fermentation is used to produce the ethanol.
Clause 38 the method of clause 30, wherein the ethanol is produced using lignocellulosic sugars.
Clause 39 the method of clause 38, wherein Simultaneous Saccharification and Fermentation (SSF) or Step Hydrolysis Fermentation (SHF) is used to produce ethanol.
Clause 40. A composition comprising the yeast strain of clause 1 or the derivative of clause 2, and one or more ingredients selected from the group consisting of surfactants, emulsifiers, gums, swelling agents, protectants, and antioxidants.
Clause 41-the composition of clause 40, wherein the composition comprises one or more defined features selected from the group consisting of: (a) Higher ethanol yields than Fali under the same fermentation conditions; (b) Has increased temperature tolerance compared to Fali at temperatures of 32 ℃ to 36 ℃; (c) Lower glycerol yields than Fali under the same fermentation conditions; (d) Having increased organic acid tolerance compared to Fali at a pH of from about 4.0 to about 5.2 in the presence of an organic acid; and (e) has an increased fermentation rate compared to Fali under the same fermentation conditions.
Clause 42. A method of producing ethanol from biomass by contacting the biomass with the composition of clause 40.
The method of clause 43, clause 42, wherein the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.
Clause 44 the method of clause 42, wherein the ethanol is produced using starch.
Clause 45 the method of clause 44, wherein Simultaneous Saccharification and Fermentation (SSF) or continuous fermentation is used to produce the ethanol.
Clause 46. The method of clause 42, wherein the ethanol is produced using sugar.
The method of clause 47, 46, wherein batch fermentation or continuous fermentation is used to produce the ethanol.
Clause 48 the method of clause 42, wherein the ethanol is produced using lignocellulosic sugars.
Clause 49 the method of clause 48, wherein the ethanol is produced using Simultaneous Saccharification and Fermentation (SSF) or Step Hydrolysis Fermentation (SHF).
Clause 50. A method of producing a fermentation product from a substrate by contacting the substrate with a fermenting organism, wherein the fermenting organism is selected from the group consisting of: (a) A yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003, or a derivative thereof; (b) A yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004, or a derivative thereof; (c) A yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005, or a derivative thereof; (d) A yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006, or a derivative thereof; (e) A yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit number Y-68007, or a derivative thereof; (f) A yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008, or a derivative thereof; and (g) a yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009, or a derivative thereof.
Clause 51 the method of clause 50, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or a combination thereof.
The method of clause 52, clause 50, wherein the fermentation product is ethanol.
Clause 53 the method of clause 52, wherein the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.
Clause 54 the method of clause 50, wherein the fermenting product is produced using batch fermentation, continuous fermentation, simultaneous Saccharification and Fermentation (SSF), or Step Hydrolysis Fermentation (SHF).
8. Preservation of organisms
Representative samples of yeast (Saccharomyces) strains as described herein were saved at the agricultural research services patent culture Collection (NRRL) at 12 months 10 of 2020, university street 1815, piolyte, illinois, U.S.A. And are designated NRRL patent deposit number Y-68003 (Y1912), NRRL patent deposit number Y-68004 (Y1913), NRRL patent deposit number Y-68005 (Y1914), NRRL patent deposit number Y-68006 (Y1919), NRRL patent deposit number Y-68007 (Y1923), NRRL patent deposit number Y-68008 (Y1927) and NRRL patent deposit number Y-68009 (Y1929) at month 23 of 2020. The preservation will be at the NRRL preservation site for at least thirty years and at least five years after the preservation site received the last request to provide a preserved sample, as specified by the international recognition of the budapest treaty on preservation of microorganisms for patent procedures. Applicant has met all of the requirements of 37c.f.r. ≡1.801-1.809, including providing proof of sample survival. Additional storage will be made on the NRRL as needed to ensure availability for the conditions described herein. After the patent is issued in this application, the applicant has no restrictions whatsoever on the usability of the NRRL deposit material. Applicant does not have the right to revoke any restrictions on the transfer of biological materials or the global commercial transport. The applicant does not relinquish any rights granted in any country in accordance with any patent issued in this application.
Claims (54)
1. A yeast (Saccharomyces) strain selected from the group consisting of:
(a) Yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003;
(b) Yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004;
(c) Yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005;
(d) Yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006;
(e) Yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit No. Y-68007;
(f) Yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008; the method comprises the steps of,
(g) A representative sample of the yeast (Saccharomyces) strain Y1929, which was deposited under NRRL patent deposit number Y-68009.
2. A derivative of a yeast (Saccharomyces) strain selected from the group consisting of:
(a) Yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003;
(b) Yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004;
(c) Yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005;
(d) Yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006;
(e) Yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit No. Y-68007;
(f) Yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008; the method comprises the steps of,
(g) A representative sample of the yeast (Saccharomyces) strain Y1929, which was deposited under NRRL patent deposit number Y-68009.
3. The yeast strain of claim 1 or the derivative of claim 2, wherein the yeast strain or the derivative comprises one or more defined features selected from the group consisting of:
(a) Under the same fermentation conditions, higher ethanol yields than yeast (Saccharomyces) strain Y1609;
(b) Has increased temperature tolerance compared to yeast (Saccharomyces) strain Y1609 at a temperature of 32℃to 36 ℃;
(c) Lower glycerol yields than yeast (Saccharomyces) strain Y1609 under the same fermentation conditions;
(d) Has increased organic acid tolerance compared to yeast (Saccharomyces) strain Y1609 in the presence of an organic acid at a pH of from about 4.0 to about 5.2; and
(e) Under the same fermentation conditions, there is an increased fermentation rate compared to yeast (Saccharomyces) strain Y1609.
4. The yeast strain or the derivative according to any one of the preceding claims, wherein the yeast strain or the derivative produces at least about 0.5% more ethanol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
5. The yeast strain or the derivative according to any one of the preceding claims, wherein the yeast strain or the derivative produces at least about 2% less glycerol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
6. The yeast strain or the derivative according to any one of the preceding claims, wherein the fermentation rate of the yeast strain or the derivative after 24 hours of fermentation is at least about 2% higher than that of saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
7. A yeast strain or the derivative according to any one of claims 3-6, wherein the temperature is 32 ℃.
8. A yeast strain or derivative according to claim 3, wherein the yeast strain or derivative produces at least about 5% more ethanol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
9. A yeast strain or derivative according to claim 3, wherein the yeast strain or derivative produces at least about 3% less glycerol after 50 hours of fermentation relative to saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
10. A yeast strain or the derivative according to claim 3, wherein the fermentation rate of the yeast strain or the derivative after 24 hours of fermentation is at least about 10% higher than that of saccharomyces cerevisiae (Saccharomyces cerevisiae) strain Y1609.
11. A yeast strain or derivative according to any one of claims 3 or 8-11, wherein the temperature is 36 ℃.
12. The yeast strain or the derivative of any one of claims 3-11, wherein the organic acid comprises lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, or a combination thereof.
13. A method of producing a derivative of the (Saccharomyces) yeast strain of claim 2 comprising:
(a) Providing:
(i) A first yeast strain, wherein the first yeast strain is selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; and
(ii) A second yeast strain, wherein the second yeast strain is in a yeast narrow-variety (Saccharomycessensu stricto) clade;
(b) Inducing sporulation of the first yeast strain and the second yeast strain;
(c) Screening and selecting spores from the first yeast strain and spores from the second yeast strain;
(d) Hybridizing selected spores of the first yeast strain with selected spores of the second yeast strain; and
(e) Screening or selecting the derivative strain.
14. The method of claim 13, wherein step (c) comprises screening or selecting spores that exhibit one or more defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.
15. The method of claim 13, wherein step (e) comprises screening or selecting hybrids that exhibit one or more defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.
16. A method of producing a derivative of the yeast (Saccharomyces) strain of claim 2, comprising:
(a) Providing:
(i) A first yeast strain, wherein the first yeast strain is selected from the group consisting of yeast (Saccharomyces) strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; and
(ii) One or more additional yeast strains in the yeast narrow (Saccharomyces sensu stricto) clade;
(b) Inducing sporulation of the first yeast strain and the one or more additional yeast strains to produce spores;
(c) Mixing all spores of step (b) to allow hybridization of the spores; and
(d) Screening or selecting the derivative strain.
17. The method of claim 16, wherein step (d) comprises screening or selecting hybrids that exhibit one or more defined characteristics of yeast (Saccharomyces) strain Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929.
18. A mutant yeast strain according to claim 1 or a derivative according to claim 2.
19. A method of producing the mutant yeast of claim 18, wherein the mutant yeast is mutated by contacting the yeast strain with a mutagen.
20. The method of claim 19, wherein the mutagen is Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine-2 (ICR-170), or nitrogen mustard.
21. A method of producing the mutant yeast of claim 18, wherein the mutant yeast is mutated by contacting the derivative with a mutagen.
22. The method of claim 21, wherein the mutagen is Ethyl Methanesulfonate (EMS), ultraviolet (UV), X-rays, methyl Methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine, 2-methoxy-6-chloro-9[3- (ethyl-2-chloroethyl) aminopropylamino ] acridine-2 (ICR-170), or nitrogen mustard.
23. An evolved yeast of the yeast strain of claim 1 or the derivative of claim 2.
24. A method of producing the evolved yeast of claim 23, wherein evolution is induced by applying selection pressure to the yeast strain.
25. A method of producing the evolved yeast of claim 23, wherein the evolution is induced by applying a selection pressure to the derivative.
26. A genetically modified yeast strain according to claim 1 or derivative according to claim 2.
27. The genetically modified yeast of claim 26, wherein the nucleic acid sequence of the genetically modified yeast is altered using genetic editing.
28. A recombinant yeast strain according to claim 1 or a derivative according to claim 2.
29. The recombinant yeast of claim 28, wherein the recombinant yeast comprises modifications to inhibit gene expression, enhance gene expression, introduce genes, or delete genes.
30. A method of producing ethanol from a substrate by contacting the substrate with a fermenting organism, wherein the fermenting organism is selected from the group consisting of:
(a) A yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003, or a derivative thereof;
(b) A yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004, or a derivative thereof;
(c) A yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005, or a derivative thereof;
(d) A yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006, or a derivative thereof;
(e) A yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit number Y-68007, or a derivative thereof;
(f) A yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008, or a derivative thereof; the method comprises the steps of,
(g) Yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009, or a derivative thereof.
31. The method of claim 30, wherein the substrate comprises or is derived from sugarcane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or combinations thereof.
32. The method of claim 30, wherein the yeast strain comprises one or more defined features selected from the group consisting of:
(a) Under the same fermentation conditions, higher ethanol yields than yeast (Saccharomyces) strain Y1609;
(b) Has increased temperature tolerance compared to yeast (Saccharomyces) strain Y1609 at a temperature of 32℃to 36 ℃;
(c) Lower glycerol yields than yeast (Saccharomyces) strain Y1609 under the same fermentation conditions;
(d) Has increased organic acid tolerance compared to yeast (Saccharomyces) strain Y1609 in the presence of an organic acid at a pH of from about 4.0 to about 5.2; and
(e) Under the same fermentation conditions, there is an increased fermentation rate compared to yeast (Saccharomyces) strain Y1609.
33. The method of claim 30, wherein the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.
34. The method of claim 30, wherein the ethanol is produced using starch.
35. The method of claim 34, wherein Simultaneous Saccharification and Fermentation (SSF) or continuous fermentation is used to produce the ethanol.
36. The method of claim 30, wherein the ethanol is produced using a sugar.
37. The method of claim 36, wherein batch fermentation or continuous fermentation is used to produce the ethanol.
38. The method of claim 30, wherein the ethanol is produced using lignocellulosic sugars.
39. The method of claim 38, wherein Simultaneous Saccharification and Fermentation (SSF) or Step Hydrolysis Fermentation (SHF) is used to produce ethanol.
40. A composition comprising the yeast strain of claim 1 or the derivative of claim 2 and one or more ingredients selected from the group consisting of surfactants, emulsifiers, gums, swelling agents, protectants and antioxidants.
41. The composition of claim 40, wherein the composition comprises one or more defined features selected from the group consisting of:
(a) Higher ethanol yields than Fali under the same fermentation conditions;
(b) Has increased temperature tolerance compared to Fali at temperatures of 32 ℃ to 36 ℃;
(c) Lower glycerol yields than Fali under the same fermentation conditions;
(d) Having increased organic acid tolerance compared to Fali at a pH of from about 4.0 to about 5.2 in the presence of an organic acid; and
(e) Under the same fermentation conditions, there is an increased fermentation rate compared to Fali.
42. A method of producing ethanol from biomass by contacting the biomass with the composition of claim 40.
43. The method of claim 42, wherein the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.
44. The method of claim 42, wherein the ethanol is produced using starch.
45. The method of claim 44, wherein Simultaneous Saccharification and Fermentation (SSF) or continuous fermentation is used to produce the ethanol.
46. The method of claim 42, wherein the ethanol is produced using a sugar.
47. The method of claim 46, wherein batch fermentation or continuous fermentation is used to produce the ethanol.
48. The method of claim 42, wherein the ethanol is produced using lignocellulosic sugars.
49. The method of claim 48, wherein Simultaneous Saccharification and Fermentation (SSF) or Step Hydrolysis Fermentation (SHF) is used to produce the ethanol.
50. A method of producing a fermentation product from a substrate by contacting the substrate with a fermenting organism, wherein the fermenting organism is selected from the group consisting of:
(a) A yeast (Saccharomyces) strain Y1912, a representative sample of which has been deposited under NRRL patent deposit number Y-68003, or a derivative thereof;
(b) A yeast (Saccharomyces) strain Y1913, a representative sample of which has been deposited under NRRL patent deposit number Y-68004, or a derivative thereof;
(c) A yeast (Saccharomyces) strain Y1914, a representative sample of which has been deposited under NRRL patent deposit number Y-68005, or a derivative thereof;
(d) A yeast (Saccharomyces) strain Y1919, a representative sample of which has been deposited under the NRRL patent deposit number Y-68006, or a derivative thereof;
(e) A yeast (Saccharomyces) strain Y1923, a representative sample of which has been deposited under NRRL patent deposit number Y-68007, or a derivative thereof;
(f) A yeast (Saccharomyces) strain Y1927, a representative sample of which has been deposited under NRRL patent deposit number Y-68008, or a derivative thereof; the method comprises the steps of,
(g) Yeast (Saccharomyces) strain Y1929, a representative sample of which has been deposited under NRRL patent deposit number Y-68009, or a derivative thereof.
51. The method of claim 50, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, tapioca, or combinations thereof.
52. A process according to claim 50, wherein the fermentation product is ethanol.
53. The method of claim 52, wherein the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.
54. The method of claim 50, wherein batch fermentation, continuous fermentation, simultaneous Saccharification Fermentation (SSF), or Step Hydrolysis Fermentation (SHF) is used to produce the fermentation product.
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| US202163170884P | 2021-04-05 | 2021-04-05 | |
| US63/170,884 | 2021-04-05 | ||
| PCT/US2022/023343 WO2022216622A1 (en) | 2021-04-05 | 2022-04-04 | Yeast strain development for ethanol production |
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| CN117897475A true CN117897475A (en) | 2024-04-16 |
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| EP (1) | EP4320220A1 (en) |
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| US20100311065A1 (en) * | 2009-06-01 | 2010-12-09 | Ubersax Jeffrey A | Genetically modified microbes producing isoprenoids |
| ES2464740T3 (en) * | 2009-06-01 | 2014-06-03 | Amyris, Inc. | Method to generate a genetically modified microbe |
| JP2014508516A (en) * | 2011-01-28 | 2014-04-10 | アミリス， インコーポレイテッド | Screening of microcolonies encapsulated in gel |
| WO2015099727A1 (en) * | 2013-12-26 | 2015-07-02 | Hill's Pet Nutrition, Inc. | High nutrient yeast |
| EP3344748B1 (en) * | 2015-09-03 | 2020-11-04 | Vib Vzw | Yeast alleles involved in tolerance to high alcohol levels |
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| JP2024515048A (en) | 2024-04-04 |
| AR125297A1 (en) | 2023-07-05 |
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| WO2022216622A1 (en) | 2022-10-13 |
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